European Conference of National Strategies for Chlamydia Trachomatis and Human Papillomavirus May 25-27, 2011, Jurmala, Latvia Baltic Beach hotel CONFERENCE BOOK Please visit Conference webpage: www.cthpv.org Auspices Conference arises from the Project European Conference of National Strategies for Chlamydia Trachomatis and Human Papillomavirus-NSCP which has received funding from the European Union, in the framework of the Public Health Programme. 1 cobas 4800 CT/NG Test cobas CT/NG Test Robust4800 CT and NG testing in a simplified workflow ® ® Robust CT and NG testing in a simplified workflow Multiple Testing Options This test is being designed to run as CT only, NG only, or as CT/NG combination. Multiple Testing Options This test is being designed to run as CT only, NG only, or as Plans include validating test for use at launch with endocervical swabs CT/NG combination. Multiple Specimen Types Plans include validating test for use at launch with endocervical swabs as well as male and female urine. Other specimen types or collection Roche proprietary collection kit cobas® PCR media to ensure devices must beroom validated by customers. at least 90 days temperature sample stability for urine and swab Multiple Specimen Types as well as male and female urine. Other specimen types or collection devices must be validated by customers. New Collection Kit for Longer Sample Stability & Primary Tube samples. The cobas® PCR media tubes can be automatically loaded Loading onto the Automated ® on to the system without transfer secondary Preparation PCRtube. media to ensure Roche proprietary collection kit to cobas NewSample Collection Kit for Instrument Longer Sample Stability & Primary Tube at least 90 days room temperature sample stability for urine and swab ® •400 µL for swab samples 10% of the available samples. The cobas PCRwhich mediaequals tubesless canthan be automatically loaded Minimal Sample Input Loading onto the Automated PCR media sample cobas®transfer on to thecollected system inwithout to secondary tube. Volume Required Instrument Sample Preparation •850 µL for urine samples which equals less than 10% of the available Minimal Sample Input Volume Required sample cobas® PCR media •400 µL collected for swabinsamples which equals less than 10% of the available ® PCRand media sample cobas Residualcollected specimenincan be stored used for repeat testing if necessary •850 µL for urine samples which equals less than 10% of the available sample collected in cobas® PCR media Higher Sensitivity for Detecting CT & NG with Automated Front End Sample Preparation Higher Sensitivity for Detecting CT & NG with Automated Front End Sample Preparation 2 Sample preparation is based on state-of-the-art magnetic glass Residual specimenThis canisbesuperior stored to and used for repeat testing particles chemistry. crude process methods by ifremoving necessary inhibitory materials and concentrating target DNA, resulting in a more robust and reliable test. Sample preparation is based on state-of-the-art magnetic glass particles chemistry. This is superior to crude process methods by removing inhibitory materials and concentrating target DNA, resulting in a more robust and reliable test. High Throughput The cobas® 4800 System is designed to process up to 384 samples per day. Less Hands-On Time Primary collection tubes can be placed directly onto the sample preparation instrument. Automation eliminates the need for manual handling of samples, reducing time and the potential for cross contamination. Manual processing intervention is limited to the transfer of the plate to the cobas z 480 analyzer. High NG Specificity Newly designed test that targets a well conserved region (DR-9) along with a specific variant sequence unique to the Neisseria gonorrhoeae species. The NG assay includes a total of 4 primers and 2 probes which amplify and detect both wild type and variant versions of this target sequence. No cross reactivity with commensal Neisseria or other bacterial species have been observed with this new NG assay. Dual target for C. trachomatis detection The cobas® 4800 CT/NG Test simultaneously detects two DNA targets; one within the C. trachomatis cryptic plasmid and the other in the CT genome. Therefore, targeting infections caused by wild type CT, the Swedish variant, or other chlamydia strains that may harbor deletions in the cryptic plasmid, as well as CT that do not carry the cryptic plasmid. Robust Combo Test Roche tests are designed to be robust and minimize the effects of PCR competition in a multi-target assay. The cobas® 4800 CT/NG Test is being designed to detect CT or NG targets within a 1:10,000 ratio of each other (10 copies of CT in the presence of 100,000 copies of NG; 10 copies of NG in the presence of 100,000 copies of CT). Quality Controls •Internal Control for full process monitoring – from extraction to detection, ensuring effective sample preparation, amplification, and target detection •Positive and Negative Controls are included in every run – to validate the results obtained •AmpErase enzymes controls contamination from amplicon carry over Clear and Definitive Results Results can be Positive, Negative, or Invalid.* No indeterminate results are expected; reduces need for retesting. * Invalid results identify specimen inhibition detected by the Internal Control as means to prevent false negative results. Workflow Timeline Total run time to result: Sample prep: Detection: Hands-on time: 4.5 hours per run – 94 patient samples 2 hours 2 hours 30 minutes (estimated)* Next run can start as soon as the PCR plate is transferred from cobas x 480 instrument. No centrifugation or incubation required. * Limited to loading of samples and reagents and transfer of microwell plate from cobas x 480 instrument to cobas z 480 analyzer. Expected Performance: Analytical Sensitivity Design goal for Limit of Detection is one (1) Inclusion-Forming Unit (IFU) per PCR for all serotypes of Chlamydia trachomatis. Design goal for Limit of Detection is ten (10) Colony-Forming Units (CFU) per PCR for all genotypes of Neisseria gonorrhoeae. 1 INVITATION . ........................................................................................... 3 ORGANIZERS . ........................................................................................ 4 REGISTRATION . ..................................................................................... 5 SCIENTIFIC AND SOCIAL PROGRAMME . ......................................... 8 SUBMITTED ABSTRACTS ................................................................... 12 2 INVITATION You are very welcome to this Conference arises from the Project European Conference of National Strategies for Chlamydia Trachomatis and Human Papillomavirus-NSCP which has received funding from the European Union, in the framework of the Public Health Programme. Target group of the participants includes laboratory specialists, clinicians, microbiologists, surveillance experts, public health specialists, healthcare economists, healthcare funders, policy makers. Objective of the Conference is evaluation and developing of Chlamydia trachomatis and HPV current control programs. Control programs aim to reduce the prevalence diseases, but this is difficult to monitor as it requires periodic population surveys of the population. However, there are many other indicators of the effectiveness which should be built into any program from the outset. At the national level, programs should monitor indicators relating to the policies and guidelines of the program, the implementation and processes, and the outcome of the program. These must be based on the specific objectives appropriate to the level of implementation. If countries move from one level of control to the next, they will need to make decisions based on a rigorous appraisal of the evidence for effectiveness, cost-effectiveness and harms. This will be assisted if countries ensure that all activities are fully evaluated and results shared with others in Europe. This way investments in programs made now will strengthen the evidence base for Chlamydia trachomatis and HPV control and facilitate future decision making and improve population health. At the European level, the objective should be to reduce the proportion of countries reporting no organized activity. Prof. Aija Zilevica, Lead microbiologist from the University of Latvia, Conference President Prof. Mario Milco D’Elios, Lead immunologist from the University of Florence, Chairperson of the Conference Scientific Committee Dzintars Ozolins, MD, PhD President of the Latvian Society of Laboratory specialists, Chairperson of the Conference Steering Committee INVITATION Purpose of the Conference is to provide guidance in the European Union about national strategies for Chlamydia trachomatis and human papillomavirus (HPV) early detection and control. It will provide a framework for developing, implementing or improving national strategies to control of Chlamydia trachomatis and HPV. Health policies, like clinical guidelines, should be based on the best available evidence. In this Conference we will aim to facilitate the development of local, evidencebased guidelines within the context of sound national strategies. Such strategies need to take account not only of clinical and epidemiological factors (such as the prevalence in the population) but also of local systems of health care delivery, infrastructure and resourcing. 3 ORGANIZERS Conference Scientific committee: Christopher Barbara (Malta), Bertille de Barbeyrac (France), Maria Jose Borrego (Portugal), Tania Crucitti (Belgium), Alexandros Daponte (Greece), Marius Domeika (Sweden), Mario Milco D’Elios (Chairperson, Italy), Viorica Gheorghiu (Romania), Karin Haar (Germany), Bjorn Herrmann (Sweden), Steen Hoffmann (Denmark), Kai Joers (Estonia), Margaretha Jurstrand (Sweden), Hilde Klovstad (Norway), Juta Kroica (Latvia), Vesta Kucinskiene (Lithuania), Catherine M Lowndes (Health Protection Agency), Servaas A. Morre (The Netherlands), Sander Ouburg (The Netherlands), Dzintars Ozolins (Latvia), Michael Pfleiderer (Germany), Mirja Puolakkainen (Finland), Dace Rezeberga (Latvia), Magdalena Rosinska (Poland), Peter Sasieni (United Kingdom), Emma Savage (Health Protection Agency), Par Sparen (Sweden), Magnus Unemo (Sweden), Inga Velicko (Sweden), Aija Zilevica (Latvia) Expert group: Mario Milco D’Elios (Chairperson, Italy), Marita van de Laar (ECDC), Catherine M Lowndes (Health Protection Agency), Dzintars Ozolins (Latvia), Magnus Unemo (Sweden) Impact and expected outcomes of the Conference will be realized by Scientific committee reviewing submitted abstracts for publishing, and providing ideas during meetings of the Conference for the Expert group. Expert group will ensure scientific control and long term impact of the Conference results realizing by publishing Conference book and Essentials of the Conference, preparing questionnaires and summarizing results from questionnaires. Scientific Committee will confirm abstracts for publishing and their usefulness for work of the committee during the Conference. This will provide scientific control of the Conference. Questions arising at the online based Forum for discussion will be forwarded to Expert group and this will ensure long term monitoring of the impact of the Conference results. Conference Steering committee: Parsla Gredzena, Violeta Mavcutko, Dzintars Ozolins (Chairperson), Vita Scepetova ORGANIZERS Steering committee is responsible for organization, planning and management of the Conference. 4 REGISTRATION Registration fees include the entrance to the Conference sessions, Conference materials, coffee brakes, lunch and Gala dinner. Registration fees do not include accommodation costs and travel expenses. Registration for accompanying person - entrance to Gala dinner - 60 EUR REGISTRATION Registration fees later 15.05.2011 just on site in 26.05.2011 - 300 EUR 5 cobas 4800 HPV Test cobas 4800 HPV Test Clinically relevant, reliable, and accurate ® ® Clinically relevant, reliable, and accurate Assay Design and Sample Options Assay Design and Sample Options Designed as a qualitative single tube multiplex assay that simultaneously detects 14 high-risk genotypes, identifies Designed a 18, qualitative single multiplex assay that HPV Type as 16 & and ß-globin is tube used as an internal control. simultaneously detects 14 high-risk genotypes, identifies Test options include: HPV Type 16 & 18, and ß-globin is used as an internal control. •HPV-HR only (which provides qualitative result) •HPV-HR + Genotype Test options include:(GT) 16/18 (which provides HPV-HR result and separate qualitative resultqualitative for Type 16 result) and Type 18) •HPV-HR only (which provides Multiple Specimen Types Multiple Specimen Types Primary Vial Loading On the Automated Sample Preparation Instrument •HPV-HR + Genotype (GT) 16/18 (which provides HPV-HR result ® TM and Liquid Plans validation in bothresult PreservCyt andinclude separate qualitative for Type 16SurePath and Type 18) Based Cytology (LBC) vials. A Roche labeled specimen collection kit is also planned for stand-alone DNA tests for samples run separately Plans include validation in both PreservCyt® and SurePathTM Liquid or cases where conventional PAP smears are common. Based Cytology (LBC) vials. A Roche labeled specimen collection kit is also planned for stand-alone DNA tests for samples run separately Sample carriers are available to load primary LBC vials directly or cases where conventional PAP smears are common. onboard the sample prep system, minimizing hands on time for transfer of sample. Primary Vial Loading On the Automated Preparation Minimal Sample Sample Input Instrument Volume Required Sample carriers are available to load primary LBC vials directly onboard prep system, minimizing on time for Only 400the µLsample input volume is required for each hands test. Depending on transfer sample. residual of volume remaining from PAP process, options include: loading Minimal Sample Input Volume Required Only 400 µL input volume is required for each test. Depending on residual volume remaining from PAP process, options include: loading Sample preparation is based on the state-of-the-art magnetic glass primary vial or transferring to a secondary tube (requires 800 µL total particles chemistry. Onboard heating and mixing of media and cervical volume). Ideal when residual samples are below 4 mL. State-of-the-Art Sample Prep Sample preparation contamination risk. is based on the state-of-the-art magnetic glass particles chemistry. Onboard heating and mixing of media and cervical ® cells provides superior preparation product. Specialized The cobas 4800 Systemsample is designed to process up to 280 samples pipetting technology combined with enzymes cross in one day. One test can provide up to 3AmpErase results: (HPV-HR, GTreduces 16, GT 18) to enhance contamination risk.testing efficiency. State-of-the-Art Sample Prep primary vial or transferring to a secondary tube (requires 800 µL total volume). Ideal when residual samples are below 4 mL. cells provides superior sample preparation product. Specialized pipetting technology combined with AmpErase enzymes reduces cross High Throughput High Throughput The cobas® 4800 System is designed to process up to 280 samples in one day. One test can provide up to 3 results: (HPV-HR, GT 16, GT 18) to enhance testing efficiency. Less Hands-On Time Collection tubes are placed directly onto the sample preparation instrument, eliminating the need for manual handling of samples, reducing time and the potential for cross contamination. Manual processing intervention is limited to the transfer of the plate to the cobas z 480 analyzer for amplification and detection. Clinical Relevance Newly designed primers and probes target a well defined L1 region for DNA Expect to correlate to detect medically relevant pre-cancer ≥ CIN2+ Medical decision point to balance clinical specificity and clinical sensitivity to ensure application is suitable for screening and to achieve guidelines and benchmarks 1 Information on infection with genotypes 16 and 18, which have been shown to increase risk for cervical disease and provide “actionable” data for immediate follow-up 2 Assay designed to correctly identify genotypes important for HPV–HR panel and not to cross react with low–risk types Robust PCR Test Quality Controls •Internal Control ( ß-globin) for full process monitoring – from extraction to detection, ensuring effective sample preparation, amplification, and target detection •Positive and Negative Controls are included in every run – to validate the results obtained •AmpErase enzymes helps control contamination from amplicon carry over Results can be Positive, Negative, or Invalid.* Clear and Definitive Results No indeterminate results are expected; reduces need for retesting. * Invalid results identify specimen inhibition detected by the Internal Control as means to prevent false negative results. Total run time to result: Sample prep: Detection: Hands-on time: Workflow Timeline 5 hours per run – 94 patient samples 2 1/2 hours 2 hours 30 minutes (estimated)* Next run can start as soon as the PCR plate is transferred from cobas x 480 instrument. No centrifugation or incubation required. * Limited to loading of samples and reagents and transfer of microwell plate from cobas x 480 instrument to cobas z 480 analyzer. 1 Meijer Chris JLM, Berkhof, J, Castle PE et al. Guidelines for human papillomavirus DNA test requirements for primary cervical screening in women over 30 years and older. Int J of Cancer 124, 516-520 (2009) 2 Khan MJ, Castle PE, Lorincz AT et al. Elevated 10 year risk of cervical precancer and cancer in women with human papillomavirus (HPV) Type 16 or 18 and the possible utility of type specific HPV testing in clinical practive. J Natl Cancer Inst. 2005:97:1072-1079 ROCHE, COBAS, LIFE NEEDS ANSWERS, and AMPERASE are trademarks of Roche. PreservCyt is a registered trademark of Hologic, Inc. SurePath is a registered trademark of Becton, Dickinson and Company © 2009 Roche Molecular Systems, Inc. Roche Molecular Diagnostics 4300 Hacienda Drive Pleasanton, CA USA 94588 http://molecular.roche.com SCIENTIFIC AND SOCIAL PROGRAMME 25.05.2011 Workshop of Scientific committee – CLOSED MEETING. Preparing Essentials of the Conference using Conference book and latest approaches from Expert side 26.05.2011 9:00 - 12:00; 10:30 - 11:00 Coffee break Plenary session (Baltic Beach hotel, room Jura + Jurmala). Survey of Chlamydia surveillance in Europe. Leading specialists are invited to present current surveillance systems in different European countries focusing on strengths as well as weaknesses. Chairpersons: Aija Zilevica (Latvia), Charlie Jennison (Health Protection Agency), Karin Haar (Germany) 9:00 - 9:10 Opening. Aija Zilevica (Latvia) 9:10 - 9:40 Charlie Jennison (Health Protection Agency) “Surveillance of Chlamydia trachomatis in England: a regional perspective” 9:40 - 9:55 Questions, discussion 9:55 -10:15 Karin Haar (Germany) “Epidemiology of Chlamydia trachomatis infections in Germany: Data from the STD-Sentinel” 10:15-10:30 Questions, discussion 11:00-11:20 Steen Hoffmann (Denmark) “Laboratory Surveillance of Chlamydia trachomatis in Denmark 1994 - 2010” 11:20-11:40 Jurijs Perevoscikovs (Latvia) „Epidemiological surveillance of Chlamydia trachomatis infection in Latvia” 11:40-12:00 Questions, discussion Satellite session (Baltic Beach hotel, room Krasts I+II). Genotyping of Chlamydia trachomatis. Chairpersons: Margaretha Jurstrand (Sweden), Bjorn Herrmann (Sweden) PROGRAMME 9:00 - 9:20 Mirja Puolakkainen (Finland) “Genotyping of Chlamydia trachomatis strains” 9:20 - 9:50 Margaretha Jurstrand (Sweden) “Detection of Chlamydia trachomatis by genetic methods in a Swedish county before and after identification of the new mutated variant” 9:50 -10:20 Bjorn Herrmann (Sweden) “Epidemiological knowledge by genotyping Chlamydia trachomatis: an overview of recent achievements” 10:20-10:30 Questions, discussion 11:00-11:30 Magnus Unemo (Sweden) “The amazing story of the Swedish new variant of Chlamydia trachomatis” 8 11:30-11:50 Kai Joers (Estonia) “Prevalence and genetic structure of Chlamydia trachomatis population in Estonia” 11:50-12:00 Questions, discussion Satellite session (Baltic Beach hotel, room Selga). Routine diagnostics of Chlamydia and HPV. What about cost-efficiency? Chairpersons: Jolande Land (The Netherlands), Tania Crucitti (Belgium) 9:00 - 9:30 Jolande Land (The Netherlands) “Chlamydia antibodies as markers for late complications in cost-effectiveness models” 9:30 - 9:50 Alexandros Daponte (Greece) “Self sampling and HPV DNA test. What is the evidence?” 9:50 -10:20 Tania Crucitti (Belgium) “Cost-effective testing for Lymphogranuloma venereum: the experience in Belgium” 10:20-10:30 Questions, discussion 11:00-11:20 Vesta Kucinskiene (Lithuania) “Pooling genital samples could be a cost beneficial approach for screening C. trachomatis in young female patients in Lithuania” 11:20-11:30 Questions, discussion 12:00 - 14:00 Lunch 14:00 - 17:00; 15:30 - 16:00 Coffee break Plenary session (Baltic Beach hotel, room Jura + Jurmala). Screening programmes of Chlamydia in Europe. The analyzing of the different screening programmes in Europe will be presented. Focusing on ways how as high response rates as possible shall be achieved. Chairpersons: Marita van de Laar (ECDC), Per-Anders Mardh (Sweden) 14:00-14:30 Marita van de Laar (ECDC) “Chlamydia control in the Europe” 14:30-15:00 Inga Velicko (Sweden) “Epidemiology and control of genital Chlamydia trachomatis infection in Sweden” PROGRAMME 15:00-15:30 Sebastian Kalwij (United Kingdom) “8 years of Chlamydia screening in England, experiences and lessons learnt” 16:00-16:30 Per-Anders Mardh (Sweden) “Epidemiological aspects on surveillance and screening” 16:30-16:50 Jorma Paavonen (Finland) “Prevention of C.trachomatis and HPV infections: Are we doing enough?” 16:50-17:00 Questions, discussion 9 Satellite session (Baltic Beach hotel, room Krasts I+II). HPV surveillance and vaccine era. Chairpersons: Michael Pfleiderer (Germany), Emma Savage (Health Protection Agency) 14:00-14:30 Emma Savage (Health Protection Agency) “HPV surveillance in the vaccine era” 14:30-14:50 Peter Sasieni (United Kingdom) “How to set up a cervical screening programme if none exists” 14:50-15:20 Michael Pfleiderer (Germany) “Current understanding of safety and efficacy of HPV vaccines licensed in the EU” 15:20-15:30 Questions, discussion 16:00-16:20 Viorica Gheorghiu (Romania) “Challanges and facts on HPV vaccination in Romania” 16:20-16:30 Questions, discussion 19:00-22:00 Social programme (Gala dinner in Baltic Beach hotel 1st floor restaurant Perle and Caviar club). Algirdas Suminskas and group “Keksi” performance 27.05.2011 10:50-11:20 Coffee break 9:00-10:50 Plenary session (Baltic Beach hotel, room Jura + Jurmala). National laboratory diagnostics programmes of Chlamydia trachomatis. Presentations are focused on suggestions how the best practices shall be implemented in every European country. Chairpersons: Mario Milco D’Elios (Italy), Marius Domeika (Sweden) 9:00- 9:30 Max Chernesky (Canada) „Screening Strategies for Chlamydia and High Risk HPV: What are the Roles for Self Collection and Amplified DNA and RNA Assays?” PROGRAMME 9:30- 9:50 Marius Domeika (Sweden) “Quality enhancements and quality assurance of laboratory diagnosis of sexually transmitted infections in Eastern Europe” 9:50-10:00 Questions, discussion 10:00-10:20 Diana Dusacka (Latvia) “Detection of Chlamydia trachomatis by molecular biological methods in Latvia” 10:20-10:30 Questions, discussion 10:30-10:40 Mario Milco D’Elios (Italy) “Actual issues from the Conference Scientific committee” 10:40-10:50 Closing remarks. Dzintars Ozolins (Latvia) 10 9:00-10:50 Sattelite session (Baltic Beach hotel, room Krasts I+II). Advances in the diagnosis of Chlamydia and HPV. Scientifically oriented session concerning development of the new laboratory methods. Chairpersons: Sven Müller-Loennies (Germany), Sander Ouburg (The Netherlands) 9:00 - 9:30 Edwin Roovers (The Netherlands) “Do Abbott RealTime CT and High Risk HPV meet the requirements for laboratory diagnosis and screening of CT and HPV?”; sponsored by Abbott Molecular 9:30 - 9:50 Sander Ouburg (The Netherlands) “The value of host genetic markers combined with CT serology in tubal pathology diagnosis” 9:50 -10:20 Frank Pieksma (Switzerland) “Cobas 4800: new developments for the detection of HPV & CT/NG”; sponsored by Roche 10:20-10:30 Questions, discussion 10:30-10:50 Sven Müller-Loennies (Germany) “Antibodies against Chlamydia Lipopolysaccharide -- Molecular Insight into Specificities and CrossReactions” 9:00-10:50 Round table (Baltic Beach hotel, room Selga). New approaches to manage Chlamydia and HPV screening. Chairpersons: Bertille de Barbeyeac (France), Catherine M Lowndes (Health Protection Agency) 9:00 - 9:20 Jolita Rimiene (Lithuania) “New methods for cervical cancer detection in Lithuania. Value of high-risk HPV-DNA testing in the triage of women with ASC category from liquid based cytology” 9:20 - 9:40 Maria José Borrego (Portugal) “Lymphogranuloma venerum recent outbreak in Europe: the Portuguese experience” 9:40 - 9:50 Questions, discussion 9:50 -10:20 Jean-Jacques Palombo (Switzerland, Roche Scientific Division) “Advances in the diagnosis of Chlamydia and HPV - what happens in industry?” PROGRAMME 10:20-10:40 Bertille de Barbeyrac (France) “Is glans swab appropriate for diagnosis of Chlamydia trachomatis infection in asymptomatic men?“ 10:40-10:50 Questions, discussion Commercial presentations and/or fairs Roche - Baltic Beach hotel, room Dzintars Abbott Molecular - Baltic Beach hotel, room Liedags Gen-Probe - Baltic Beach hotel, room Banga BD - Baltic Beach hotel, room Krasts III 11 SUBMITTED ABSTRACTS IS GLANS SWAB APPROPRIATE FOR DIAGNOSIS OF C.TRACHOMATIS INFECTION IN ASYMPTOMATIC MEN? Corresponding author: Bertille de Barbeyrac Institution of corresponding author: Université de Bordeaux, INRA, USC Mycoplasmal and chlamydial infections in humans, French National Reference Center for chlamydial infections, 33076 Bordeaux, France Country: France E-mail: [email protected] With nucleic acid amplified tests (NAATs) that are sensitive and specific for detection of C. trachomatis, non-invasive specimens such as first catch urine (FCU) and self-collected vaginal swabs can be used. While FCU is the alternative to the urethral swabs, specimen transport can be messy and problems have been associated with the handling of FCU in the laboratory. Swabs are a better specimen for processing. A few studies have shown that self-collected glans swab may be appropriate for C. trachomatis NAATS. We evaluated self-collected glans swab vs FCU in asymptomatic heterosexual men and MSM attending the screening center (anonymous and free of charge) in Bordeaux, France [1]. A subject first self-collected penile swab using a flocked regular swab (Copan Italia Spa, Italy, Mast Diagnostic, France) and after self-collected urine. The flocked swab was transported dry to the laboratory and transferred into M4RT transport medium (500µl) (method 1) or into urine (500µl) (method2). Both samples were simultaneously tested for C. trachomatis using the PCR assay (Cobas TaqMan CT Test, Roche Diagnostic). The load of C. trachomatis was estimated by using cycle thresholds (Cts values) given by Cobas TaqMan in all positive specimens. ABSTRACTS For 344 men included in method1, the sensitivity was 89% (24/27) for urines and 67% (18/27) for penile swabs. The comparison of means of Cts values showed a difference of at least 4 Cts between urine (33.2) and penile swab (38.5) when both were positive or when only one was positive, urine (38) and penile swab (42). For 259 men included in method 2 (pooling strategy), the sensitivity of 94.7% was identical for urines and for penile swabs discharged in urine. The comparison of means of Cts values showed similar Cts between urine (35.4) and penile swab in urine (36.7). When only one test was positive, Cts were high, 41.2 for urines and 40.7 for penile swabs. In asymptomatic men, the number of bacteria is often very low and could explain – the discrepancies of both specimens, - the low sensitivity of penile swab, - the non reproducibility of results. Our results agree with those of two others studies [2, 3]. These results suggest the self-collected penile swab may not be useful for C. trachomatis testing. The penile swab could not replace FVU in screening programs 1 Raherison S, Peuchant O, Clerc M, et al. Glans swabs are not appropriate specimens for diagnosis of Chlamydia trachomatis infection in asymptomatic men. J Clin Microbiol. 2009; 47: 2686. 12 2 Moncada J, Schachter J, Liska S, Shayevich C, Klausner JD. Evaluation of self-collected glans and rectal swabs from men who have sex with men for detection of Chlamydia trachomatis and neisseria gonorrhoeae by use of nucleic acid amplification tests. J Clin Microbiol. 2009; 47: 1657-1662. 3 Pittaras TE, Papaparaskevas J, Houhoula DP, et al. Comparison of penile skin swab with intra-urethral swab and first void urine for polymerase chain reaction-based diagnosis of Chlamydia trachomatis urethritis in male patients. Sex Transm Dis. 2008; 35: 999-1001. LYMPHOGRANULOMA VENEREUM RECENT OUTBREAK IN EUROPE: THE PORTUGUESE EXPERIENCE Corresponding author: Maria José Borrego Other authors: Gomes JP* Nunes A* Florindo C* Borges V* Ferreira R* Santo I** Azevedo J** Institution of corresponding author: National Institute of Health, Lisbon, Portugal Institutions of other authors: *National Institute of Health, Portugal **STD clinic, Lapa Health Center, Lisbon, Portugal Speciality: Molecular microbiology Country: Portugal E-mail: [email protected] Introduction: In Portugal no surveillance measures were launched as a consequence of the 2004 lympgogranuloma venereum (LGV) outbreak in Europe. However, all Chlamydia trachomatis specimens that are sent to the National Institute of Health (NIH) of Portugal are systematically genotyped. This procedure potentially enables the opportunistic detection of Portuguese circulating LGV strains. Objectives: To report the unusual number of LGV specimens observed during the 20072010 period, and to analyze clinical and epidemiologic data of LGV infected individuals. Additionally, to compare the outer membrane protein A gene (ompA) sequence of LGV specimens with both L2-434 reference strain and L2-variants described in the literature. Results: So far, a total of 514/833 (61,7%) C. trachomatis specimens could be ompA genotyped and 17/514 (3,3%) matched LGV-genotypes (16 "L2" and 1 mixed E+L2 undetermined variant). LGV-genotypes were observed in both men (n=10; 5 MSM+1 heterosexual+4 unknown; 4 presented proctitis of which 2 were HIV+) and women (n=7; all heterosexual, 6 asymptomatic and 1 PID). All the LGV samples detected in 2007 revealed ompA sequences different from both L2-434, and L2b-144276. LGV specimens detected during 2008 and 2010 were similar to L2b-144276, while the ones detected during 2009 were similar to the L2 prototype strain. 13 ABSTRACTS Methods: During the 2007-2010 period, C. trachomatis infection was tested over 9000 samples by the Cobas-Amplicor (2007-2009) or the Cobas 4800 (2010), yielding 833 (9,2%) positive results. DNA was extracted from C. trachomatis-positive samples, using the QIAamp DNA Mini Kit. Amplification and automated-sequencing of the ompA gene, using Laser-Gene99 software (DNASTAR), were used for genotyping. Discussion: From 1991 to 2002 only 5/463 typed specimens were L2 strains. During 2002-2006 no LGV strain was detected. So, the recent number of L2 specimens was quite unexpected, probably related with the recent worldwide outbreak of LGV. During 2007 two studies (1 Portuguese, 1 Austrian) reported new L2 variants, but no link could be established with specific clinical findings. Globally, the majority of the amino acid changes on the L2 variants occur in a well-defined antigenic domain. Conclusions: The particular location of the mutations of the newly described L2 variants suggests a chlamydial strategy for host immune evasion. The lack of surveillance systems for C. trachomatis infections, including LGV, in Portugal, makes this outbreak has passed completely unnoticed and prevented the taking of any control measures or to establish any link between the Portuguese LGV cases and European (or other) sexual networks. KNOWLEDGE, ATTITUDES AND BEHAVIOUR ABOUT SEXUALLY TRANSMITTED DISEASES (STDs): A SURVEY AMONG ITALIAN UNIVERSITY FEMALE STUDENTS Corresponding author: Chiara Cadeddu Other authors: Chiara de Waure*, Maria Rosaria Gualano*, Alice Mannocci°, Giacomina Chiaradia^, Daniela Vincitorio°°, Francesco di Stanislao°°, Elisabetta De Vito**, Elisa Langiano**, Antonio Boccia°, Walter Ricciardi*, Giuseppe La Torre° Institution of corresponding author: Hygiene Institute - Catholic University of Sacred Heart (Rome, Italy) Institutions of other authors: *Institute of Hygiene, Catholic University of the Sacred Heart, Rome, Italy °Clinical Medicine and Public Health Unit, Sapienza University of Rome, Rome, Italy ^Spallanzani Institute, Rome, Italy °°Section of Hygiene and Public Health, Polytechnic University of the Marche Region, Italy **Department of Motor Sciences and Health, Local Health Agency, Cassino, Italy Speciality: Hygiene & Preventive Medicine Country: Italy ABSTRACTS E-mail: [email protected] Background: Sexually Transmitted Diseases (STDs) represent a considerable Public Health issue. Although in the second half of the last century the diffusion of effective screening and prevention tools has made possible important improvements in terms of mortality, there is still a great concern about STDs and young women appear as the most affected by the “development deficit” of the potential of STDs prevention. Objectives: To find out the level of knowledge and attitudes about STDs and preventive measures among young women and deepen the understanding of variables associated to them. 14 Methods: We designed a questionnaire accounting mostly for knowledge about STDs and knowledge and attitudes toward Pap test and vaccination against Human Papilloma Virus (HPV), and submitted it to 313 young female university students (mean age: 22 ys), opportunistically identified. Chi-square test and Mann-Whitney test were performed in order to identify associated factors. Results: Knowledge about STDs seems to be weak, in contrast to knowledge about preventive measures, which appears good but limited. About STDs ways of transmission, 85.3% of women who answered that the use of condom and intercourses with a single known partner are effective behaviours for preventing STDs showed a right knowledge of STDs ways of transmission in comparison to 67.2% of women giving incorrect answers (p<0.001). Eighty three point two per cent of women who took information about STDs from Internet, books and TV showed a correct knowledge of STDs ways of transmission, in comparison to 67.3% (p=0.002) among the remainder. For Pap test the following results came up: 100% of women with a high level of knowledge on STDs knew Pap test in contrast to less than 90% among the remainder (p=0.04); moreover 89.4% of the women that were aware of effective behaviours for preventing STDs (condom and intercourses with a single known partner) knew the Pap test in comparison to 80.8% of the women that were unaware of them (p=0.04). Conclusions: Women are still not completely aware of risks for STDs and preventive measures to avoid them. Informational and educational campaigns should be organised to reach this target and limit the current and future burden of STDs. REVIEW OF GENITAL CHLAMYDIOSIS AND RISK FACTORS IN LITHUANIA IN 2007 - 2009 Corresponding author: Caplinskiene Irma Institution of corresponding author: Lithuanian Centre for Communicable Diseases and AIDS, Mykolas Romeris University Speciality: Medical epidemiologist, MD Country: Lithuania Introduction: The incidence of Chlamydia trachomatis in Lithuania was to 9.6 cases per 100000 persons in 2009, and in comparison with 2006 (11.9 cases per 100000 persons) it has decreased. There were 326 cases of Chlamydia infection registered in 2009, and 403 cases in 2007 and 2008 in Lithuania. Though, it is believed that the incidence of Chlamydia infection is substantially higher than that officially recorded. Objectives: To assess the trends of Chlamydia trachomatis incidence and related risk factors (behavioral and sociodemographic) over the last 3 years (2007-2009) in Lithuania. Methods: Statistical analysis of data (sociodemographic characteristics and sexual risk behavioral factors) of Chlamydia trachomatis infection cases through the period of 2007-2009. The data was accumulated by the National Centre for Communicable Diseases and AIDS. 15 ABSTRACTS E-mail: [email protected] Results: Trends: steady increase of chlamydia cases among 20-29 year old male who had 1-3 female sex partners; the number of chlamydia cases among those who indicated a casual partner as a likely source of infection increased; however, the number of chlamydia cases among those who could not tell the likely source of infection decreased. There were fewer cases of infection among individuals who reported constant use of condoms, whereas the number of chlamydia cases among those who reported occasional use or non-use of condoms increased; no differences were found in terms of education, place of residence, age group, social insurance, social groups, and disease detection place, compared to 2007, 2008 and 2009. Conclusions: There is an annual increase of genital chlamydia infections in the group of young males from urban areas. There were fewer cases among individuals who reported constant use of condoms. No differences were found in terms of education, place of residence, age group, social insurance, social groups, disease detection place, and whether or not the person was engaged in a commercial sex scenario, compared to 2007, 2008 and 2009. The contact tracing deteriorates with each year. SCREENING STRATEGIES FOR CHLAMYDIA AND HIGH RISK HPV: WHAT ARE THE ROLES FOR SELF-COLLECTION AND DNA AND RNA ASSAYS? Corresponding author: Max Chernesky, PhD Institution of corresponding author: McMaster University/St. Joseph’s Healthcare, Hamilton, ON, Canada Country: Canada Introduction: Sexually transmitted Chlamydia trachomatis (CT) and human papillomaviruses (HPV) are very common worldwide. Because the majority of these infections are asymptomatic, screening programmes using non-invasive sampling methods are required to identify and treat lower genital tract CT infections to prevent ascending upper tract complications. The screening goal for HPV testing is to test women for high risk HPV adjunctively to cervical Pap testing or vaginal testing of women who decline pelvic examination, in order to identify and treat women with precancerous lesions. Sensitive and specific nucleic acid assays are available for these purposes. ABSTRACTS Objectives: To review comparative data on the use of commercial assays for CT and high risk (HR) HPV using cervical swabs and Pap collections versus vaginal and urine specimens in women and urethral, urine, penile meatal, anal and oral swabs in men. Methods: Publications from the peer-reviewed literature and recent data from the author’s laboratory will be presented to provide evidence for using various specimen types and DNA and rRNA assays for CT diagnosis. The performance of DNA and E6/E7 mRNA assays will be reviewed for HR HPV diagnosis. Results: CT infections are most prevalent in sexually active, younger women and men. Lower genital tract asymptomatic infection rates can be as high as 75% allowing inapparent ascending infections in women and unknowing transmission of infection between sexual partners. Specimens of choice for screening are self-collected vaginal swabs for women, and first catch urine and/or self-collected penile swabs for men. There is good evidence for 16 screening self-collected anal swabs from men who have sex with men. Although women over 30 are primary candidates for Pap and HPV testing, in some societies Pap tests are being performed on younger women. These specimens can serve as samples of convenience for screening for CT. Several commercial molecular tests for CT are being used and in comparative studies the results from these assays may be impacted by analytical sensitivity, stability of nucleic acids in specimens, and inhibitors. Comparisons of different collection devices and manipulation of specimens in the preanalytical phase of testing may influence testing outcomes. Similar studies for HPV have shown that most specimen types contain DNA and E6/E7 mRNA. Most of the commercial assays have comparative sensitivities for detecting CIN2+ pathology and have been shown to facilitate colposcopy examinations for patients with ASC-US cytology. The role for HPV adjunctive testing and screening in select populations is becoming clearer. Discussion: Sampling, testing, automation and focused screening will be discussed with an emphasis on research gaps. Conclusions: Manufacturers of clinically relevant diagnostic assays for CT and HPV deserve credit for advancing our knowledge of the role of these pathogens in patient morbidity. Screening programmes should use these assays wisely. CHLAMYDIA SCREENING IN IRELAND - NOT READY YET? RESULTS OF A PILOT STUDY OF OPPORTUNISTIC SCREENING FOR GENITAL CHLAMYDIA TRACHOMATIS INFECTION IN THE REPUBLIC OF IRELAND Corresponding author: Dr. Emer O' Connell 1 Other authors: D Vaughan 2, R Brugha 3, M Balfe 3, M Cormican 2,4, P Gillespie 2, C O' Neill 2, C Coleman 4, C Fleming 2,4, M Fitzgerald 1, A Murphy 2, D O'Donovan 2,5. Institution of corresponding author: 1Health Service Executive (HSE) Dublin/Mid-Leinster, Ireland Institutions of other authors: 1 Health Service Executive (HSE) Dublin/Mid-Leinster, Ireland 2 National University Ireland, Galway, Ireland 3 Royal College of Surgeons, Dublin, Ireland 4 Galway University Hospitals, Ireland 5 Health Service Executive (HSE) West, Galway, Ireland Speciality: Public Health Medicine Country: Republic of Ireland Introduction: Chlamydia trachomatis (CT) is an infection of public health concern in the Republic of Ireland with annual notifications rising from 245 in 1995 to 1,278 in 2003. In 2004, legislation requiring laboratory notification came into effect, and the number increased to 6,290 in 2008 [1]. CT is now the most commonly notified sexually transmitted infection (STI) in Ireland. Irish prevalence data shows asymptomatic rates of 3.7-11.2 for females and 5.9% for males. There is no formal organized screening programme for CT in the Republic of Ireland. Objectives: A chlamydia screening pilot study was conducted in Ireland between 2007 and 2009 to assess the feasibility and cost effectiveness of introducing a screening programme 17 ABSTRACTS E-mail: [email protected] for CT in the Republic of Ireland. 18-29 years olds were the eligible population. 16 and 17 year olds were excluded because of ethical restrictions under Irish law; and the range 18 to 29 years includes the highest notification group (20-29 years old) in Ireland. Methods: Opportunistic screening was offered to women and men aged 18-29 years who attended urban and rural general practices, student health units, and a family planning clinic in the west of Ireland. Non clinical 'pee-in-a-pot' (PIP) days were organized in two higher education institutions. Urine specimens were tested with polymerase chain reaction technology (COBAS TAQMAN). Economical evaluation compared the pilot strategy of opportunistic screening in terms of costs, including those averted by screening, and effectiveness. Results: 998 eligible people were tested: 460 in clinical settings and 538 in non clinical PIP settings. Screening offer rates were low (1-9%), refusal rates were low, and urine testing was highly acceptable. 48 people (4.8%, 95% CI: 3.5-6.1) tested CT positive. Management of cases, partner notification and outcome rates corresponded well with best international practice. Screening in the combined clinical settings would result in an incremental cost per quality adjusted life year (QALY) gained of 94,717 EUR, while 'pee-in-a-pot' screening was estimated to result in 42,967 EUR per QALY gained. Discussion: A research health adviser was essential for coordinating the care of positive cases, especially for partner notification. Low offer rates in clinical settings reflected the time constraints on General Practitioners and practice nurses. Opportunistic screening in the combined healthcare settings is not cost-effective. Neither screening settings are considered cost-effective. Conclusions: We recommend prioritizing primary prevention and the provision of supports to primary care providers for optimal case diagnosis (where clinically indicated), management and follow-up. Dedicated regional health advisers could provide support and advice to clinicians and cases, and coordinate partner notification, testing and treatment. References: (1) HPSC. The Health Protection Surveillance Centre. Sexually Transmitted Infections 2008. Annual summary report. 2010. COST-EFFECTIVE TESTING FOR LYMPHOGRANULOMA VENEREUM: THE EXPERIENCE IN BELGIUM Corresponding author: Tania Crucitti ABSTRACTS Other authors: Hilde Smet Institution of corresponding author: Institute of Tropical Medicine Institutions of other authors: Institute of Tropical Medicine Speciality: Microbiology Country: Belgium E-mail: [email protected] 18 Introduction: Lymphogranuloma venereum is reported in several countries in Europe. It has been accepted that the infection is confined to men having sex with men usually presenting symptoms of proctitis. The testing for L genotypes of Chlamydia trachomatis is very often restricted to this high risk population. Up to date laboratories rely on in-house methods to identify the L genotypes which can be costly and time consuming depending on the method used. Objectives: To apply a nucleic acid amplification testing algorithm to firstly screen specimens for C. trachomatis and to secondly identify the non-L and L genotypes without increasing considerably the cost of testing, hands-on time and turnaround time. Methods: C. trachomatis positive DNA extracts tested with the Abbott CT/NG Real Time PCR were re-tested using an in-house Real Time-PCR (RT-PCR) assay to determine whether the C. trachomatis belonged to the L genotypes. The RT-PCR differentiated L from non L genotypes. As part of the validation of the in-house RT-PCR its performance was compared to the Restriction Fragment Length Polymorphism (RFLP) assay used so far. The algorithm was validated on urine, genital, pharyngeal and anal specimens. Results: The Abbott CT/NG Real Time PCR and in-house RT-PCR showed identical sensitivities and specificities for genital, anal, pharyngeal and urine specimens. Fifty two positive anal specimens were tested in the RFLP and RT-PCR assay. Both methods identified 8 non-L, 42 L and 1 mixed non-L and L type infection. One specimen was a non-L type by RT-PCR but undetermined using RFLP. Since the introduction of the algorithm in September 2010 and till February 2011, a total of 1124 specimens were tested; 66 were C. trachomatis positive with 53 non-L and 13 L genotypes. All results were reported within 2 days after specimen reception. The routine use of the RT-PCR assay increased the cost with only 10%. Conclusions: We identified L genotypes of C. trachomatis at an affordable cost, within a reasonable time frame, and in all C. trachomatis positive specimens. By consequence we are now able to identify LGV infection in the general population or in patients in which LGV infection was not suspected. At the end the patients will benefit from a more timely and reliable diagnosis and appropriate management. Finally we will gain insight into the LGV epidemiology. 19 ABSTRACTS Discussion: The presented algorithm improved the diagnosis of LGV considerably. The L genotypes were identified immediately after obtaining a C. trachomatis positive result and independently from the origin of the specimen and risk behavior of the patient. This is in contrast with the previously used RFLP method which was performed on batches of specimens and on only anal specimens collected from MSM. In addition, the hands-on time for the in-house RT-PCR method was reduced by using the same DNA extract as the screening assay. SELF SAMPLING AND HPV DNA TEST. WHAT IS THE EVIDENCE? Corresponding author: Alexandros Daponte Country: Greece E-mail: [email protected] Background: During the last decade, increasing efforts have focused on increasing the overall proportion of women participating in cervical cancer screening procedures, mainly by HPV detection in self-obtained samples, such as vaginal and urine specimens as reported by our team previously (Daponte et al. 2003, 2004). HPV screening during antenatal visits may increase participation, facilitate early detection and be cost-effective for patients and the health services. Objectives: A study of screening high-risk (hr) human papillomavirus (HPV) types in first void urine and vaginal samples in pregnant women attending the antenatal clinic using our optimized protocol for PCR detection in urine and vaginal samples in order to establish the feasibility of using these specimens for HPV detection and compare them with non pregnant women hr HPV incidence rates from the same area in central Greece (University hospital of Larissa). Study design: Paired urine and vaginal specimens obtained from 500 pregnant women attending antenatal clinic were included. Vaginal swabs and urine samples were tested in all participants for HPV types 16, 18, 31 and 45 after urine microscopy was performed. Prevalence of HPV was compared with 500 paired urine and vaginal specimens from non pregnant women collected by midwives in the same area. ABSTRACTS Methods: Urinalysis and urine microscopy was performed. QIAamp Viral RNA Mini Kit (Qiagen Inc., Valencia, CA) was used for DNA extraction. An in house PCR method was used for amplifying the above hr HPV types. The quality of sample was validated by detection of the housekeeping beta-globin gene. Results: The hr HPV detection rate was similar in pregnant and non pregnant women. In the antenatal patients, 15/500 paired urine/vaginal samples were both positive for the same type of hrHPV. Only one sample was vaginal positive/urine negative. The urine/vaginal detection sensitivity was higher in pregnancy because first void urine samples from pregnant women usually contain two or more epithelial cells per field in urine microscopy increasing HPV detection as reported by our team before. Conclusions: Urine and vaginal samples are easily collected during antenatal visits and may be particularly important in screening young women who are asymptomatic and for personal or cultural reasons would not participate in a cervical specimen screening program. This study indicates that urine and vaginal specimens in pregnancy are easily obtainable and adequate for screening for hr HPV but this should be confirmed in a larger future prospective trial. 20 CHLAMYDIA IN SWEDEN - KEY FACTORS OF SUCCESSFUL PREVENTIVE RESPONSE Corresponding author: Charlotte Deogan Other authors: Cecilia Moberg, PhD Anna Månsdotter, PhD Lene Lindberg, PhD, Associate Professor Institution of corresponding author: Department of Public Health Sciences, Karolinska Institutet, Stockholm Sweden. Institutions of other authors: Department of Public Health Sciences, Karolinska Institutet, Stockholm Sweden Speciality: Public health Country: Sweden E-mail: [email protected] Introduction: After a continuous increase of Chlamydia trachomatis in Sweden, a general reduction in the incidence was seen in 2009. Incidence and prevention activities varied largely between geographical regions. Objectives: The aim of the study was to identify key factors of successful regional prevention of chlamydia and other sexually transmitted infection (STIs) by a mapping of the preventive response in seven Swedish counties and a case comparison related to chlamydia incidence. Methods: A multiple case study was performed including seven Swedish counties based on data from surveys and structured interviews with key stakeholders, county council registry data and national surveillance data on chlamydia incidence and testing. Factors of prevention were rated in a case comparison across counties resulting in a matrix of prevention factors. Key factors of successful prevention were identified by contrasting high incidence counties with low incidence counties. Discussion: Results were found to be compliant with earlier research. Further research and evaluation is however needed of specific measures to explore what constitutes an effective prevention mix. Conclusions: Successful prevention of chlamydia at the regional level requires strong leadership and coordination of preventive measures that incorporates multiple collaboration partners and research knowledge. Adequate resource investments in both primary- and secondary prevention and a broad mix of prevention activities with high testing coverage targeting risk individuals are key factors in successful preventive health care response. 21 ABSTRACTS Results: Counties with a reduction in chlamydia incidence apply adequate investments in STI-prevention, simultaneously implement a broad mix of activities, involve multiple local agents, include efforts to target risk individuals with testing and counseling and use the internet as a platform for prevention activities. Other key factors identified were a strong leadership, managing regional networks and being strongly connected to research. Furthermore, the combination of the above-mentioned activities, together with regional participation in national planning and strategy for chlamydia prevention, could be a vital part of the prevention mix. QUALITY ENHANCEMENTS AND QUALITY ASSURANCE OF LABORATORY DIAGNOSIS OF SEXUALLY TRANSMITTED INFECTIONS IN EASTERN EUROPE M. Domeika,1 R Ballard,2 and M Unemo3 on behalf of the Eastern European Network for Sexual and Reproductive Health (EE SRH Network) Department of Medical Sciences, Uppsala University, Uppsala, Sweden ([email protected]) 2 Division of STD Prevention, Centers for Disease Control and Prevention (CDC), Atlanta, USA 3 Swedish Reference Laboratory for Pathogenic Neisseria, Department of Laboratory Medicine, Clinical Microbiology, Örebro University Hospital, Örebro, Sweden 1 Background. Sexually transmitted infections (STIs), excluding HIV, are not considered as major burden of public health in Eastern European countries and therefore are left aside, e.g. mainly no national strategies or programs exist, and laboratory diagnosis, patient management and epidemiological surveillance are suboptimal. Initial goal: To optimize, harmonize and quality assure the laboratory diagnosis of STIs in the catchment countries. Methods. Through participation in the multinational and multidisciplinary Eastern European Network for Sexual and Reproductive Health (EE SRH Network), presently including active representatives from 14 EE countries as well as international STI experts from EU countries and USA, to reach consensus on ideal performance and logistics for the laboratory diagnosis of STIs with further implementation and legalization of the materials elaborated on country level. ABSTRACTS Results. Following consensus has been reached: sole use of microscopy for the diagnosis of N gonorrhoeae should be limited to the diagnosis of male symptomatic urethritis and use of culture and NAATs have to be promoted; surveillance of antimicrobial resistance is crucial to establish. NAATs have to become the method of choice for the diagnosis of C trachomatis and M genitalium, and wider implemented for diagnosis of T vaginalis, syphilis and herpes infections. Serology for diagnosis of C trachomatis and T vaginalis infections has to be abolished. Wet smear microscopy is still recognized as a sufficient tool for the diagnosis of T vaginalis and bacterial vaginosis (BV). BV can also be diagnosed using a Gram-stained smear. NAATs that are not internationally acknowledged should be validated against the international standard assays. International and nationally adjusted guidelines have been produced and published (English and respective national languages). Conclusion. Adoption of internationally acknowledged, evidence-based standards will allow countries improvement of their medical care standards. Since its formation, the EE SRH Network has been effective in facilitating this process. 22 DETECTION OF CHLAMYDIA TRACHOMATIS BY MOLECULAR BIOLOGICAL METHODS IN LATVIA Corresponding author: Diana Dusacka Other authors: Aija Gulbe, Natalia Repuscenko, Natalia Novikova, Violeta Mavcutko, Tatjana Kolupajeva, Jelena Storozenko Institution of corresponding author: State Agency "Infectology Center of Latvia"( ICL) Institutions of other authors: State Agency "Infectology Center of Latvia"( ICL) Speciality: microbiologist Country: Latvia E-mail: [email protected] Introduction: Chlamydia trachomatis is a mandatory notifiable infection in Latvia. Different diagnostic methods for detection of C. trachomatis were used in Latvia. In early 1990s Chlamydia serology and Chlamydia antigen detection assays - DFA (Direct Fluorescent antibody test) and EIA (Enzyme Immune Assay) were widely used. Since 1996s some laboratories started Chlamydia detection by DNA probe assay and Nucleic Acid Amplification Technique (NAAT). Objectives: The aim of this study is to give short overview of methods, currently used in Latvia and to analyze experience of testing by molecular biological technologies. Methods: Data from the national computer-based surveillance system VISUMS and ICL laboratory data were used to analyze laboratory methods for confirmation of Chlamydia infection in Latvia. Discussion: According to published data, NAAT are the most sensitive (90-95%) tests for genital Chlamydia (DFA sensitivity: 65-75%). In spite of this fact, in Latvia NAAT is not widely used. Possible reasons are: expensive equipment and reagents, special rooms requirements, need for high personal skills. New technology validation has to be continued, however preliminary ICL data showed approximately the same rate of positive results by DFA and NAAT in a limited group of samples. DFA results can be very sensitive due to experience of microscopist, but the method is not suitable for large workloads. Conclusions: 474/1042 (45.5%) cases of Chlamydia infection registered in 2010 were confirmed by molecular biological methods, but only 9.4% - by NAAT. High sensitivity and specificity of NAAT tests, simultaneous detection of several agents (e.g. C. trachomatis and N. gonorrhoeae), possibility to use non invasive clinical samples for testing, make it suitable for improvement of C.trachomatis diagnosis, and for large volume screening. 23 ABSTRACTS Results: 1042 laboratory confirmed cases of Chlamydia infection were registered in Latvia in 2010: 329 (31.6%) positive results were identified using DFA, 378 (36.3%) - DNA probe assay, 96 (9.4%) - NAAT. C.trachomatis DFA method traditionally was mostly used in ICL laboratory. In 2010, 3829 and 126 patients were tested by DFA and by DNA Probe assay, respectively. Frequency of positive results for C. trachomatis was 5% and 6.3%, respectively. A NAAT based method (Aptima Combo 2 Assay Gene-Probe), was recently introduced in ICL. Parallel testing of clinical specimens (swabs, urine) from 100 patients were performed by both methods: 6/100 were positive by DFA and 7/100 by NAAT. PREVALENCE OF HIGH-RISK HUMAN PAPILLOMAVIRUS INFECTION IN CERVIX CYTOLOGIC SMEARS IN ALBANIA Corresponding author: Kozeta Filipi1 Other authors: Federico De Marco2, Gentian Kusta3, Mirela Rista4, Albana Ahmeti1. Institution of corresponding author: 1. Institute of Public Health, Department of Epidemiology, Tirana, Albania Institutions of other authors: 2. Laboratory of Virology – “Regina Elena” Cancer Institute, Rome, Italy 3. Maternity Hospital, Berat, Albania 4. Maternity Hospital, Tirana, Albania Speciality: epidemiologist Country: Albania E-mail: [email protected] Introduction: Cervical cancer ranks the 2nd most frequent cancer in women in Albania, and the 2nd most frequent cancer among women between 15 and 44 years of age (IARC, GLOBOCAN 2008). The established cause is high-risk types of human papillomavirus (HPV), and a new modality for cervical cancer screening is the combination of cervical cytology with HPV testing. Objectives: To identify the prevalence of HPV infection, the types of HPV and cervical cytological profiles of healthy Albanian women. Methods: This cross-sectional investigation of HPV typing together with cytology screening in Albania was conducted between 2004 and March 2007, covering a total of 402 women. HPV detection and typing were carried out by a combined MY09/MY11 and GP5+/GP6+ PCR followed by direct sequencing of generated amplicons. ABSTRACTS Results: The mean age of the study group was 36 years (range 19-75 years) and the prevalence of high risk HPV infection was 15%. This figure was higher (25.2%) among women aged between 19-29 and lower (13.6%) among those aged 30 or over. HPV 16 was found to be the most frequent type (41% of cases), followed by HPV 53 (7.2%), HPV 31 (5.8%), and HPV 18 (4.3%). HPV 81 and HPV 84 were the most prevalent low-risk types detected with prevalence of 11.6% and 5.8%, respectively. Discussion: The use of a simple HPV DNA test followed by immediate ‘screen and treat’ algorithms based on visual inspection in those who are HPV positive are needed to minimize the number of visits and make best use of limited resources. Detailed knowledge of HPV type circulating patterns in specific local geographical areas is essential for appropriate implementation of screening, prevention, and surveillance campaigns. Conclusions: In this study no differences were noted in types of HPV between young and mature women. The circulation of HPV types is far more complex than assumed generally. 24 CHALLENGES AND FACTS ON HPV VACCINATION IN ROMANIA Corresponding author: Viorica Gheorghiu 1* Other authors: Aurora Stanescu, Alina Zaharia, Denisa Janta, Teodora Solomon 1* Institution of corresponding author: National Center for Surveillance and Control of Communicable Diseases 1* Institutions of other authors: National Center for Surveillance and Control of Communicable Diseases 1* Speciality: public health and epidemiology Country: Romania E-mail: [email protected] Introduction: Despite no specific serological survey regarding the prevalence of the HPV carriers being conducted in the last 10 years, the decision makers and those responsible from the Ministry of Health have decided to introduce the HPV vaccination in November 2008. The target group was girls aged12-13 years. The implementation of this vaccination campaign had many constraints. Objectives: Identification of knowledge, attitude and behavior which led to acceptance or rejection of the 2 types of HPV vaccines (Cervarix and Silgard) by children from the target group or/and their tutors. Methods: The National Center for Information and Prevention of Cervix Cancer organized with the help of National Center for Surveillance and Control of Communicable Diseases an information campaign between 15.05.2009 - 18.12.2009 using a "green line" phone and an electronic information site. Discussion: After the implementation of information campaign the acceptability of the HPV vaccine improved but still there are many who refused the second or third doses. For avoiding the wastage of the vaccine the population target group was extended up to 24 years of age. The HPV vaccination campaign had a negative impact on routine vaccination too. Conclusions: The introduction of the new vaccines is more and more difficult to be accepted by general population and the information campaign for HPV vaccine should to precede and start at least 4 months before vaccination itself. 25 ABSTRACTS Results: During the six months of the campaign a total of 3626 phone calls and 362 emails were recorded. From this huge number, only 1955 calls were specific for cervical cancer and HPV infection. All the questions have been coded and then analyzed. The main topics of the questions were focused on: information regarding the development and implementation of the campaign (17,57%), information about the vaccines used (33,79%), information regarding cervical cancer (48,69%). EPIDEMIOLOGY OF CHLAMYDIA TRACHOMATIS INFECTIONS IN GERMANY: DATA FROM STD-SENTINEL Corresponding author: Karin Haar Haar K (1), Bremer V (2), Sailer A (1), Marcus U (1), Hamouda O (1) (1) Department for Infectious Disease Epidemiology, HIV/AIDS, STI and Bloodborne Infections Unit, Robert Koch-Institute, Berlin, Germany (2) EPIET Chief coordinator, European Centre for Disease Prevention and Control (ECDC), Stockholm, Sweden Country: Germany E-mail: [email protected] Introduction: Chlamydia trachomatis (CT) infections are often asymptomatic, especially in women, and can lead to infertility. In Germany, the new Infection Protection Act was implemented in 2001 and only syphilis, HIV and hepatitis were stated reportable STIs. A STD sentinel surveillance system was set up by the end of 2002, where a subset of health care providers voluntarily reported the incidence of STIs. Further, in 2008 an opportunistic chlamydia screening programme for all women under the age of 25 was introduced in Germany. Objectives: To collect data on the frequency of STIs, the demographic features and geographic distribution, a clinical STD-sentinel surveillance system was implemented in a country where most STIs are not reportable. We tried to identify risk groups and behaviour to improve guidance on targeted prevention and future surveillance initiatives. Methods: Between the end of 2002 and 2009 local health authorities, specialized outpatient clinics and practitioners continually reported examination and infection data, demographic characteristics and possible risk factors for STI-acquisition. Patients were asked to return completed questionnaires which were linked to doctors’ reports and provided further information about patients’ social state, sexual behaviour and possible source of infection. ABSTRACTS Results: Of all chlamydia-examinations, 6% (5955/98405) were positive. This was higher than for any other lab-confirmed STI, namely gonorrhoea (GO) 3.6%, HIV 1.1%, syphilis 3% and trichomonas 2.5%. For 2640 chlamydia cases (64% women) further data were provided: The overall mean age was 26 years, men being significantly older. 63% of women were migrants, 70% sex workers and 45% of men were MSM. 13% of women had CT in the past, 17% of men had a previous episode of GO. In total, 10% were co-infected with GO. 54% of women stated “health check” as their reason of attendance, whereas 23% reported “health problems”. 35% of women believed their regular partner to be the source of infection, 33% named customers, 61% of men reported casual partners. The median number of sexual partners within the last 6 months was 3 in men and 2 in women. 23% of men and 47% of women reported consistent, 32% of men and 6% of women reported no condom use with casual partners. 26 Discussion: Due to high rates of CT-infected women who only attended for a “health check” i.e. were asymptomatic, CT might be an underdiagnosed but frequent infection in Germany. For countries with lacking STI-notification systems, implementation of an STD-sentinel seems feasible; however, data have to be interpreted cautiously as selection bias is always an issue. Patients seen in STD-Sentinel institutions have high re- and co-infection rates, are frequently working in the sex industry and are migrants or MSM. Therefore data from the sentinel cannot be interpreted as infection rates in the general population. Conclusion: Clinical STD-sentinel surveillance systems are useful to match laboratory findings with behavioural data; however, recruitment sites have to be chosen according to the studied core group. The German CT screening programme is now accompanied by CT-laboratory sentinel surveillance. Data from this lab sentinel will provide more reliable public health data on the epidemiology of chlamydia in Germany. EPIDEMIOLOGICAL KNOWLEDGE BY GENOTYPING CHLAMYDIA TRACHOMATIS: AN OVERVIEW OF RECENT ACHIEVEMENTS Corresponding author: Björn Herrmann Institution of corresponding author: Department of clinical microbiology, Uppsala University Hospital, Sweden Country: Sweden Email: [email protected] Overview: Several C. trachomatis typing methods based on other target genes than ompA have been published in recent years. Two multilocus sequence typing (MLST) schemes based on housekeeping genes have been shown useful for analysis of evolutionary changes and slowly evolving processes [1, 2]. Their differentiating capacity is similar to ompA sequence analysis and therefore they have limitations in applications for short term epidemiogy. A third MLST system was developed by Klint et al. [3] and is intended for clinical epidemiology and outbreak investigations. It is based on five highly variable but stable genomic loci which give the system up to a fivefold higher resolution than ompA. A variety of clinical specimens have been analysed, including urogenital chlamydia [4-6], LGV [7] and trachoma [8]. There is also a multilocus variable number tandem repeat (VNTR) analysis (MLVA) system which offers a resolution similar to that of the MLST system by Klint et al. [9, 10]. However, both MLST and MLVA require several days of work until final results are achieved and they are expensive. Diagnostic DNA microarray technology has emerged as an alternative in microbial genotyping and reduces the cost and the work load. An ompA array has been published [11], but still no high resolution array for C. trachomatis. 27 ABSTRACTS Background: Chlamydia trachomatis is one of the most common sexually transmitted infections world-wide. Typing of C. trachomatis is important to understand its epidemiology and thereby decrease the prevalence. Conventional genotyping has been predominated by analysis of the major outer membrane protein or the encoding ompA gene. However, the differentiating capacity of ompA analysis is limited and methods with higher resolution are needed to perform adequate short term epidemiological analyses. Summary: Genotyping methods for C. trachomatis have been improved in recent years and conventional ompA sequencing has limited use compared to current alternatives. References: 1. Pannekoek et al. Multi locus sequence typing of Chlamydiales: clonal groupings within the obligate intracellular bacteria Chlamydia trachomatis. BMC Microbiology 2008;8:42. 2. Dean et al. Predicting phenotype and emerging strains among Chlamydia trachomatis infections. Emerg Infect Dis. 2009;15:1385-94. 3. Klint et al. High-resolution genotyping of Chlamydia trachomatis strains by multilocus sequence analysis. J Clin Microbiol 2007;45:1410-4. 4. Herrmann et al. Emergence and spread of Chlamydia trachomatis variant, Sweden. Emerg Infect Dis 2008;14:1462-5. 5. Jurstrand et al. Characterisation of Chlamydia trachomatis by ompA sequencing and multilocus sequence typing in a Swedish county before and after identification of the new variant. Sex Transm Infect 2010;86:56-60. 6. Christerson et al. Multilocus Sequence Typing of Urogenital Chlamydia trachomatis From Patients With Different Degrees of Clinical Symptoms. Sex Transm Dis 2011; epub Feb 4. 7. Christerson et al. Typing of lymphogranuloma venereum Chlamydia trachomatis strains. Emerg Infect Dis 2010;16:1777-9. 8. Harding-Esch et al. Multi-locus sequence typing: a useful tool for trachoma molecular epidemiology. In: Stary et al (eds). Chlamydial infections Proceedings of the 12th International Symposium on Human Chlamydial Infections; Salzburg 2010. p. 55-8. 9. Pedersen et al. Highly discriminative genotyping of Chlamydia trachomatis using omp1 and a set of variable number tandem repeats. Clin Microbiol Infect 2008; 14:644-52. 10. Wang et al. Evaluation of a High Resolution Genotyping Method for Chlamydia trachomatis Using Routine Clinical Samples. PloS one 2011, 6(2):e16971. 11. Ruettger et al. Genotyping of Chlamydia trachomatis strains from culture and clinical samples using an ompA-based DNA microarray assay. Molecular and cellular probes 2010;25:19-27. LABORATORY SURVEILLANCE OF CHLAMYDIA TRACHOMATIS IN DENMARK 1994-2010 Corresponding author: Steen Hoffmann Institution of corresponding author: Statens Serum Institut, Artillerivej 5 DK-2300 Copenhagen S ABSTRACTS Speciality: Clinical microbiology Country: Denmark E-mail: [email protected] Introduction: The Danish laboratory surveillance system for oculo-genital infections caused by Chlamydia trachomatis (CT) was introduced in 1988 and became mandatory in 1994. This presentation will give an overview of the design of the surveillance system as well as some of the results, and future perspectives and challenges. 28 Objectives: The objective of the surveillance system is to monitor the occurrence of laboratory diagnosed chlamydia in Denmark. Methods: Diagnosis of CT in Denmark is performed at 15 public laboratories, 13 in clinical microbiology and two in clinical biochemistry. They use five different laboratory data systems. The laboratories report quarterly the number of tests performed and the number of diagnosed patient cases. For each case, the following variables are reported (anonymously): age, gender, specimen date, anatomical site, type of health care provider, and laboratory method used. There is no clinical reporting. Results: From 1994 to 2009 the annual number of analyses increased from approximately 270,000 to nearly 350,000 and the number of laboratory diagnosed cases of chlamydia increased from 14,000 to 30,000. In 2009, the incidence among males was 414 per 100,000 and it was 665 among females; 83% and 90%, respectively, were 15-29 years old. The incidence among females aged 18 years appears to be increasing. The fraction of males among all diagnosed cases has slowly increased from 23% to 38%. Almost 95% of all specimens were obtained by general practitioners. Since 2003 NAATs were used in more than 99% of cases. Urine is increasingly being used as specimen material, especially among males. Discussion: The strength of this surveillance system is that it is mandatory and nationwide. The weakness is that no data are reported regarding persons with negative tests. Thus, it is not possible to account for the examined population, e.g. in terms of age, gender, anatomy or indication for testing. However, a nationwide Microbiology Data Bank is currently being developed to receive real-time complete data for all positive and negative microbiology laboratory test results, including chlamydia tests. Conclusions: This surveillance system provides reliable information about the number of laboratory confirmed cases of chlamydia. As of 2007, the responsibility for interventions to reduce chlamydia was transferred from the counties (n=15) to the municipalities (n=101). This has increased the need for more detailed information about the geographical distribution of chlamydia. Also, data about the age distribution of those examined must be established in order to allocate resources more cost-effectively. SURVEILLANCE OF CHLAMYDIA TRACHOMATIS IN ENGLAND: A REGIONAL PERSPECTIVE Corresponding author: Charlie Jennison Other authors: Lynsey Emmett Iain Roddick Adrian Deas Sam Bracebridge ABSTRACTS Institution of corresponding author: Health Protection Agency Institutions of other authors: Health Protection Agency Speciality: Chlamydia surveillance Country: England E-mail: [email protected] 29 Introduction: In England genital C. trachomatis testing can be broken into three main categories. Testing that occurs in: Genitourinary Medicine (GUM) clinics, the National Chlamydia Screening Programme (NCSP) and "Community testing" outside these two specific services. GUM and NCSP data are collected as well defined disaggregate datasets, with community testing as aggregate data by most regions (6/9). In the East of England, disaggregate community testing data have been collected centrally since 2008. Objectives: To collect a disaggregate community testing dataset for the East of England and consequently combine this with the other available datasets for the East of England, covering a time period of 30 months for the 15-24 year age group. This would allow for further investigation into chlamydia testing in the region. The combined dataset would be queried to assess progress towards achieving testing levels needed for control. Methods: Data extracts from the Genitourinary Medicine Clinic Activity Dataset (GUMCAD), the NCSP database and the East of England community testing database were combined in SQL Server 2005 and then analysed using STATA (version 11). Results: Results from Jennison et al. (Journal of Public Health, 2011), will be presented along with results from the further developmental analyses. Discussion: Combining and standardising the three datasets was possible and has resulted in the most complete surveillance data of C. trachomatis testing in England currently available. There are several benefits of this dataset, for example it allows for the easy answering of commisioners' questions or the breakdown of testing by; date of attendance, geographic location, age and sex. The benefits and limitations will be expanded upon in this presentation. Conclusions: Recent improvements in data capture for chlamydia testing in England have resulted in excellent data sources for further epidemiological analyses. In combining two of these datasets enhanced surveillance reports for the East of England have already been used to inform service provision and help commissioners meet Chlamydia testing coverage targets. The dataset produced in this work takes this a step further and can be used to look in depth at a more complete picture of chlamydia epidemiology. PREVALENCE AND GENETIC STRUCTURE OF CHLAMYDIA TRACHOMATIS IN ESTONIA Corresponding author: Kai Joers Institution of corresponding author: Quattromed ABSTRACTS Country: Estonia E-mail: [email protected] Chlamydia trachomatis is one of the most common of pathogens causing sexually transmitted diseases. In the lecture I describe the prevalence of C. trachomatis in Estonia and its population structure based on samples originating from gynecologists and andrologists. The general prevalence of C. trachomatis among Estonian population is 7,3 %, based on analysis of 1200 samples. 30 The most abundant C. trachomatis serotypes in Estonia were serotype E (32,6 %), F (17,4 %), K (15,2 %), G (13,0 %), D (8,7 %), J (6,5), B (4,3 %) and H (2,2 %). This is very similar to serotype prevalence in Sweden. As one third of all samples belonged to serotype E, serotyping only has not enough discriminatory power to separate possibly relevant different strains. For better characterization of Estonian C. trachomatis population genotyping method based on Multi Locus Sequence Typing (MLST) was introduced. It was found that MLST based on 8 loci had a discriminatory index D = 0.97 and thus suitable for genotyping for clinical use. 15 unique genotypes were identified among 21 clinical samples, indicating the existence of heterogeneous population with probably long history. PREDICTION ON TUBAL FACTOR INFERTILITY BY MEASURING C. TRACHOMATIS AND CHLAMYDIAL HEAT SHOCK PROTEIN 60-SPECIFIC ANTIBODY AND CELL-MEDIATED RESPONSES – A PROSPECTIVE STUDY Corresponding author: Päivi Joki-Korpela Other authors: Tiina Rantsi, MD Christel Hyden-Granskog, PhD Jorma Paavonen, MD, PhD, Prof Tiina Rantsi, MD Heljä-Marja Surcel, PhD Aila Tiitinen, MD, PhD, Prof Hanna Öhman, MSc Institution of corresponding author: Department of Obstetrics and Gynecology, Helsinki University Central Hospital Institutions of other authors: Department of Obstetrics and Gynecology, Helsinki University Central Hospital Institute for National Health and Welfare, Oulu Speciality: Gynecology and obstetrics Country: Finland E-mail: [email protected] Introduction: Infertility is a common problem, and examinations and treatments for patients are costly. Approximately one third of infertile couples suffer from tubal factor infertility (TFI). Non-invasive methods for detecting TFI are needed for effective and economical planning of the treatment. Our group has shown retrospectively that TFI prediction model could be improved by combining four distinct markers for humoral and cell-mediated immunity. Methods: C.trachomatis and CHSP60-specific immunoglobulin G plasma antibodies have been analyzed by ELISA. Antibodies against CHSP-60 were measured using recombinant CHSP60 as an antigen. Cell mediated immunity has been studied by using peripheral blood mononuclear cells. C.trachomatis EB; serovars E and F, and recombinant CHSP 60 have been used as antigens. Results: So far, we have collected and analyzed blood samples from approximately 225 couples (450 patients), and it seems that approximately 9% of patients (4.5% couples, 2.4% man only, 1.8% woman only) are positive with these markers. We are collecting data on 31 ABSTRACTS Objectives: The aim is to validate the developed TFI prediction model in order to develop sensitive non-invasive test to be able to efficiently and economically plan further treatment for subfertile couples. patients (results on examinations and treatments, age, previous pregnancies, STDs, illnesses, medication, BMI, smoking, use of alcohol) and will analyze and compare the obtained data with results on chlamydial immune response. Conclusions: Subfertility can be caused by several factors e.g. male factors, endometriosis, hormonal factors, TFI and unexplained infertility. Laparoscopy is a golden standard when detecting TFI. However, it is invasive, costly and can cause complications to patients. Moreover, it's unable to tell the essential function of the ovarian tubes. Non-invasive methods are needed to identify TFI for effective and economical planning of the treatment. DETECTION OF CHLAMYDIA TRACHOMATIS BY GENETIC METHODS IN A SWEDISH COUNTY BEFORE AND AFTER IDENTIFICATION OF THE NEW MUTATED VARIANT Corresponding author: Margaretha Jurstrand Other authors: Björn Herrmann (BH), Hans Fredlund (HF), Magnus Unemo (MU) Institution of corresponding author: Clinical Research Centre, Örebro University Hospital, SE-70185 Örebro, Sweden Institutions of other authors: Department of Clinical Microbiology, University Hospital, SE-751 85 Uppsala, Sweden.(BH) Department of Laboratory Medicine, Clinical Microbiology, Örebro University Hospital, SE-70185 Örebro, Sweden. (HF, MU) Speciality: PhD, Laboratory Manager Country: Sweden E-mail: [email protected] Introduction: Despite mandatory partner notification in Sweden legislated in 1988, the incidence of Chlamydia trachomatis has increased each year since 1997. In 2006 a new variant of Chlamydia trachomatis (nvCT), with a deletion in the cryptic plasmid, was reported in Sweden. This deletion included the targets for the genetic diagnostic systems used in many clinical laboratories and resulted in thousands of false negative results. ABSTRACTS Objectives: To study the distribution of C. trachomatis omp1 genotypes within sexual networks and to determine if genotyping would improve partner notification. The aim was also to characterize genetic variation among wild genotype E isolates and among nvCT isolates using a high-resolution multi locus sequence typing (MLST) system. Methods: A PCR method was developed for characterization of clinical C. trachomatis isolates by sequence analysis of the omp1 gene. The method was used for characterization of genotypes in consecutive Chlamydia tissue culture positive samples from late 2006, and was compared with the results of a one-year study performed from 1999 to 2000 in the same area. Further on we used a high-resolution multi locus sequence typing (MLST) system for analysis of C. trachomatis genotype E isolates found in 2006. Results: Sequence analysis of the omp1 gene revealed that the most prevalent genotypes was C. trachomatis genotype E, followed by F, D, K, Ba, G, H, I, and J in 1999-2000. In the largest network comprising 26 individuals three different C. trachomatis genotypes were found. The distribution of genotypes was similar in year 2006 but a significant increase 32 of genotype E from 47% in year 1999-2000 to 69% in 2006 was detected. Complete MLST sequence types (STs) were obtained from 24 of 28 wild type C. trachomatis genotype E isolates and among these 14 (58%) unique STs were identified. In these 24 isolates four omp1 variants were detected. In contrast all nvCT isolates (n=41) displayed an identical ST. Conclusions: Genotyping was shown to be useful in a partner notification context, but was found to provide limited discrimination especially for the predominating genotype E. Using the MLST system the discrimination of C. trachomatis genotype E wild type isolates is considerable higher compared to omp1 sequencing, which enables improved studies in molecular epidemiology. All nvCT in 2006 was of identical genotype E which strongly indicate a clonal spread of the nvCT. EVALUATION OF CHLAMYDIA SCREENING IN DIFFERENT COMMUNITY SCREENING VENUES IN LAMBETH, LONDON, UK 2005-2010 Corresponding author: Dr Sebastian Kalwij Other authors: Rumbi Mugezi Institution of corresponding author: Amersham Vale Practice, London, UK Institutions of other authors: Lambeth and Southwark Chlamydia Screening Office Speciality: Primary Care Country: England E-mail: [email protected] Introduction: Lambeth Primary Care Trust in London was one of the first PCT’s in England, to offer screening to young patients (age 16-25), in a variety of community settings. The programme is now in its seventh year. It is well established and screens large numbers of young people each year. We tried to map the screening activity and identify screening venues who screen the most and reach those most at risk. Methods: Most screening venues used the two main laboratories at Kings College Hospital and St Thomas Hospital. Tests requested through the website (www.checkurself.org.uk) were sent to a private laboratory (TDL, The Doctors Laboratory) in Central London. Each venue has a so-called site-code and were grouped as follows: general practice, community reproductive and sexual health clinics, Brook Advisory (Charity aimed at young people, contraception and STI’s) pharmacies, midwives, outreach services and the website. These figures were collected on a quarterly basis every year since 2005. This study excluded invalid screens and all tests from Genito-Urinary Hospital Clinics as stipulated by the National Chlamydia Screening Programme. Results: 33, 317 screens where processed over 5 years (1/4/2005-31/3/2010) at the three laboratories and the results collected by the local Chlamydia Screening Office. The results were broken down by screening venues; General practice 15928 (47.8%), Community 33 ABSTRACTS Objectives: The objective is to find out which community screening venue contributes the most in both volume and positivity rate. This is an important question to answer in times of economic uncertainty and health care budget cuts. Reproductive and Sexual Health Clinics 7235 (21.7%), Brook Advisory 6911 (20.7%), Pharmacies 773 (2.3%) and midwives, outreach programmes and the website contributed 2470 (7.4%). During the same period the number of tests for chlamydia across all venues increased from 3745 tests in 2005/06 to 10.390 tests in 2009/10. Positivity rates were as follows: General Practice 7.1%, Community Reproductive and Sexual Health Clinics 11.7%, Brook Advisory 5.3%, Pharmacies 6.7%, Midwives 6.3%, Website: 4.8%. Overall positivity rate for 2009/2010 was 7.9%. Discussion: Whilst chlamydia prevalence remains high (6-9% in certain inner city areas) it is important to identify a practical approach to screen large number of asymptomatic young people year after year. Whilst certain technological innovations like internet testing sound promising, it may not replace the traditional face-to-face consultation with a trusted Health Care Professional in either Community Reproductive and Sexual Health Clinics or General Practice yet. Conclusions: In Lambeth PCT, General Practices and the Community Reproductive and Sexual Health Clinics were responsible for the vast majority of Chlamydia tests. This demonstrates that both service providers are vital in offering the Chlamydia screening service. To set up and sustain a screening programme it is important to offer opportunistic screening via venues such as general practices and Community Reproductive and Sexual Health Clinics as they see large number of young people coming through their doors and detect those most at risk. 8 YEARS OF CHLAMYDIA SCREENING IN ENGLAND; EXPERIENCES AND LESSONS LEARNT Corresponding author: Dr Sebastian Kalwij Institution of corresponding author: Amersham Vale Practice, New Cross, London. Speciality: Primary Care Country: United Kingdom E-mail: [email protected] Introduction: This presentation aims to give an overview of the National Chlamydia Screening Programme in England, which is now in its 8th year. The programme has been set up as an opportunistic screening programme and chlamydia screening is offered to all sexually active young people between the ages of 15 and 24 years in all 152 Primary Care Trusts in England. ABSTRACTS Objectives: To identify a successful strategy to screen 35% of 15-24 year olds in England. Methods: I looked at data collected by the Health Protection Agency from 2003 to 2011. Results from all laboratories participating in the programme and the Chlamydia Screening Offices are collected by the Health Protection Agency on a quarterly basis. These figures get published in the annual reports. Results: The number of tests has increased from 18.529 in 2003 to 1.2 million in 2010/11 (22.1% coverage). The most successful screening venues in terms of volume (n=975.989) are: Outreach: 23.1%, Community Sexual Health Clinics: 21.7%, General Practice: 16.5%. 34 Alternative settings like internet testing: 2.7% and testing in local pharmacies 2.2%, reached far fewer people. In terms of positivity rate, the picture looks different. Community Sexual Health Clinics: 8.0%, Internet testing 7.8%, Pharmacy: 7.1%, General Practice: 5.5% and Outreach: 2.9%. Discussion: In order to roll out a Chlamydia Screening Programme it is important to identify a successful strategy to reach high risk populations. Whilst established clinics and primary care services are already seeing many young people, novel approaches are needed to reach those who don't visit these health care facilities on a regular basis. Conclusions: It is feasible to screen a large percentage of the target population per year. A diverse approach is needed to reach those at high risk for chlamydia whilst covering a high percentage of the target population in order to make an impact on controlling Chlamydia infection and reducing its prevalence. DEVELOPMENT OF AN OMPA PYROSEQUENCING ASSAY FOR CHLAMYDIA TRACHOMATIS BASIC GENOTYPING DIRECTLY FROM UROGENITAL AND OCULAR SPECIMENS Corresponding author: Rok Kogoj Other authors: Potocnik Marko, Darja Kese Institution of corresponding author: University of Ljubljana, Medical Faculty, Institute of Microbiology and Immunology, Zaloska 4, 1000 Ljubljana, Slovenia Institutions of other authors: Potocnik M.: University Medical Centre Ljubljana, department of Dermatovenereology, Zaloska 2, 1000 Ljubljana, Slovenia; Kese D.: University of Ljubljana, Medical Faculty, Institute of Microbiology and Immunology, Zaloska 4, 1000 Ljubljana, Slovenia Speciality: Intracellular bacteria Country: Slovenia E-mail: [email protected] Objectives: The aims of this study were to develop and validate a procedure for the genetic characterization of clinical C. trachomatis strains by a newly developed pyrosequencing assay that targets the variable domains VD I and VD IV of the ompA gene and to define the prevalence of particular genotypes among symptomatic Slovenian sexually active adults. Methods: A combination of three pyrosequencing assays that cover the whole VD-I and a part of the VD-IV was optimised with eleven reference C. trachomatis strains (D-L3) and validated by comparison to ompA Sanger sequencing. Additionally, 74 urethral swabs from men, 78 cervical swabs from women and 31 ocular swabs, which were previously diagnosed as C. trachomatis positive, were evaluated by pyrosequencing, with their results compared to the results obtained by Sanger sequencing. 35 ABSTRACTS Introduction: In epidemiological studies, Chlamydia trachomatis is mainly genotyped by conventional sequencing of the ompA gene which requires long turnaround times, lots of hands on time and is relatively high cost. To our knowledge, pyrosequencing, a bioluminimetric »de novo« sequencing technique, has not yet been tested as a tool for C. trachomatis genotyping. Results: OmpA VD-I forward, VD-I reverse and VD-IV pyrosequencing products were obtained from all reference C. trachomatis strains and all patients' samples that were positive in the respective pyrosequencing PCR. After software analysis VD-I forward, VD-I reverse and VD-IV pyrosequencing products showed average "good quality" lengths of 74, 74 and 80 nucleotides respectively and a 100% match to Sanger sequences. The most prevalent genotype was E (53.4%) followed by F (19.9%), D (7.5%), K (6.8%), G (6.2%), H (3.1%), J (1.9%) and Ia & Ja (0.6%). Discussion: Pyrosequencing the ompA gene could be used as a tool for genotyping C. trachomatis in epidemiological purposes. The main concern still remains its inability to produce more than maximally 100bp sequence reads. However, when sequencing carefully selected key areas of a gene, it is possible to achieve satisfactory amounts of sequence data to differentiate between genotypes. In our study population genotype E was shown to be dominant which is in concordance with several studies all over the world. Conclusions: Pyrosequencing is a rapid and easy to perform method; it is economically effective and could be used as an alternative to conventional sequencing for C. trachomatis epidemiological purposes. Our study contributes to the creation of an informative C. trachomatis genotyping protocol, which could enable rapid strain identification and facilitate C. trachomatis genotypes distribution studies in different patients groups. POOLING GENITAL SAMPLES COULD BE A COST BENEFICIAL APPROACH FOR SCREENING CHLAMYDIA TRACHOMATIS IN YOUNG FEMALE PATIENTS IN LITHUANIA Corresponding author: Vesta Kucinskiene1 Other authors: Skaidra Valiukeviciene1, Marius Domeika2 Institution of the corresponding author: 1Department of Skin and Venereal Diseases, Medical Academy, Lithuanian University of Health Sciences, Kaunas, Lithuania. Institution of other authors: 2Department of Medical Sciences, Uppsala University, Uppsala/ Eastern European Committee of Swedish Health Care Community, Stockholm, Sweden. Speciality: Dermatovenereologist Country: Lithuania ABSTRACTS E-mail: [email protected] Introduction: Laboratory diagnostic tests to detect C. trachomatis are expensive and not accessible for every young person in Lithuania. That is one of the reasons why the sample size of testing for C. trachomatis is very low in Lithuania though epidemiological studies show the infection to be prevalent, eg. the prevalence among 15-19-year old sexually active lithuanian adolescent girls is 18.2%. Objectives: to estimate the prevalence of Chlamydia trachomatis infection among young women (20-24 year old); to estimate the accuracy of pooling vaginal samples; to compare the costs of DNA hybridization method to detect C. trachomatis, when analyzing individual and pooled vaginal samples. 36 Methods: A total of 795 female students aged 20-24 from Kaunas high schools were invited to participate in the study. The response rate was 67 %. Study participants were surveyed using a standardized questionnaire, and collected the vaginal swabs themselves following the written instructions. All vaginal samples were prepared and analyzed using Digene Hybrid Capture II CT/NG Test in accordance with the instructions of the manufacturer. Specimens were tested individually and in thoroughly mixed pools comprising 3 consecutive samples. For validation of positive results, all positive individual samples and pools were retested. According to the test results, the sensitivity, specificity and cost savings of pooling samples were assessed. Results: The analysis of the findings showed that the prevalence of C. trachomatis infection among young women 7.1%. Two-thirds (60 %) of infected C. trachomatis female students had no specific clinical signs. Multivariable analysis showed that C. trachomatis infection was twofold more common (OR – 2.37; CI 1.06; 5.3) among students who had specific STIs’ symptoms compared to those without such symptoms. The prevalence of C. trachomatis infection was threefold higher among students who had 2–3 sexual partners compared to those who had only one sexual partner (OR – 3.17; CI 1.03; 9.7). For the estimation of the population prevalence, the costs of the present pooling strategy (177 pools) were reduced by 85 % compared with the analysis of all 533 samples. The costs of the present pooling strategy with subsequent individual testing of all samples in each positive pool (177 pools and 87 individual samples from positive pools) for the estimation of the population prevalence as well as the diagnosis of each individual sample were reduced by 70%. Discussion: In order to save the reproductive health of young women, and money, in Lithuania there could be a suggestion to politicians to implement new extensive screening strategies, i.e. to organize centralized high-quality analysis of samples which can be taken non-invasively; the detection of C. trachomatis should be performed in specialized regional laboratories using pooled samples. Screening could be more cost-effective when risk factors and specific clinical symptoms related to C. trachomatis infection are discovered. Conclusions: the prevalence among young women was detected almost the same as in the other European countries; pooling vaginal samples for nucleic acid detection of C. trachomatis is an accurate and cost saving approach for screening to compare with testing of individual samples. CHLAMYDIA CONTROL IN THE EUROPE Marita van de Laar, European Centre for Disease Prevention and Control, Stockholm, Sweden Background In 2007 a survey was carried out in EU/EFTA Member States with respect to prevention and control of Chlamydia trachomatis infections. The report “Screening for Chlamydia Review in Europe” outlines the basic requirements for guidelines, diagnostics, case management, screening and surveillance. Guidance for Chlamydia control needed to be developed with the purpose to lay down the scientific basis to support Member States to strengthen and improve Chlamydia control. Target audiences are national programme managers on sexual health and STI, policy makers at (inter)national level and experts in the field. 37 ABSTRACTS Objectives To present the latest surveillance data on Chlamydia and the ECDC guidance on Chlamydia control in the Europe. Methods A technical expert group on Chlamydia control in the Member States was established. Comments from the ECDC’s Advisory Forum to the draft guidance were taken into account. The 1990-2009 surveillance data have been collected in TESSy in 2010. Results The guidance recommends a stepwise approach to Chlamydia control to ensure that prevention and patient management are in place before complex interventions such as screening are considered. A: Primary prevention (health promotion/education, school programmes, condom distribution); B: Case management (routine case surveillance, diagnostics, patient and partner management); C: Opportunistic testing (testing routinely offered to specified group(s) of people attending clinical services); D: Screening programme (organised provision of chlamydia testing to a substantial proportion of a defined population). The first step to a comprehensive control programme is the adoption of a national chlamydia control strategy based on consultation with key stakeholders. The strategy should take into account the specific national opportunities and limitations together with a review of the evidence. The strategy can be based on the stepwise approach outlined in the ECDC guidance on Chlamydia control. Nearly 344,000 cases of chlamydia have been reported in 2009; rates and trends will be presented over two decades. Conclusions The implementation of the Chlamydia guidance needs to be evaluated at both national and the EU level. At the national level, programmes should monitor indicators relating to the policies and guidelines of the programme, the implementation and processes, and the outcome of the programme. At the European level, the target is to reduce the proportion of countries reporting that no organised control activity exists. THE ROLE OF CHLAMYDIA TRACHOMATIS ANTIBODY TESTING IN THE FIRST SCREENING FOR TUBAL FACTOR INFERTILITY Corresponding author: Jaroslavs Lakutins Other authors: V.Fodina, J.Brikune Institution of corresponding author: AVA Clinic, Vilandes 3, Riga, Latvia Institutions of other authors: AVA Clinic, Vilandes 3, Riga, Latvia Speciality: gynaecologist Country: Latvia ABSTRACTS E-mail: [email protected] Introduction: Chlamydia trachomatis (CT) in the female genital tract (FGT) usually remains asymptomatic, undiagnosed and untreated. The main clinical form of CT infection is acute cervicitis. CT may ascend to the upper FGT and can cause pelvic inflammatory disease (PID) in up to 30% of cases. It increases the risk of tubal epithelial damage and tubal obstruction. Therefore, an earlier CT infection can be the reason for tubal infertility (TI). The risk of TI after the previous PID is estimated from 10 to 20%. Objectives: The association between CT IgG antibodies, which still can be in the blood many years after the treatment and tubal pathology, has been recognized before. Therefore, CT 38 antibody testing (CAT) can be used as a screening test for previous infection and possible tubal pathology on the first visit to a gynecologist in the fertility clinic. The gynecologist should separate patients with a high risk of tubal pathology for the next step of the treatment from the patients with low risk. Methods: The primary selection includes identification of the risk factors for tubal pathology during the history taking, abnormalities during the ultrasound examination (USG) and CAT. Laparoscopy is recommended for patients with known co-morbidities (PID, previous ectopic pregnancy or endometriosis), and for patients with pathology after USG. In the case of patients with no known co-morbidities and without abnormal USG finding, CAT helps to detect women with low and high risk of tubal pathology. Results: CAT has rather high negative predictive value (85-90%). It means, that the patients with negative CT antibodies (low-risk patients) have low probability of tubal damage (less than 15%). Performing hysterosalpingography (HSG) after CAT does not significantly change the probability of tubal damage. However HSG as a method has a high specificity and can very precisely confirm the absence of tubal pathology. Where appropriate expertise is available, screening for tubal occlusion using hysterosalpingo-contrast-ultrasonography (HyCoSo) should be considered because it is an effective alternative to HSG for women who are not known to have co-morbidities. Conclusions: Patients with positive CT antibodies (high-risk patients) have 53-60% probability of tubal damage (positive predictive value). HSG recognise the tubal patology with approximately the same probability. However, the risk of complications after HSG for these patients is increased by up to 10%. Therefore HSG is relatively contra-indicated for high-risk patients. Laparoscopy can be recommended to provide a final diagnosis. CT infection should be excluded before operation for high-risk patients. SHOULD WE ROUTINELY TEST MEN FOR CHLAMYDIA INFECTION IN ASSESMENT OF THEIR FERTILITY POTENTIAL? Corresponding author: Jaroslavs Lakutins Other authors: O.Lakutina Institution of corresponding author: AVA Clinic Institutions of other authors: AURA-R Medical SIA Speciality: gynaecologist E-mail: [email protected] Introduction: A history of genital tract infections (GTI) occurs in 1.6-10.3% of men attending fertility clinics. Colonization of the male genital tract is probably common and usually self-limiting. The male accessory sex glands often harbor microorganisms (also Ureaplasma urealythicum and Chlamydia trachomatis (CT)), which may colonize the urogenital tract without obvious sign of infection. 39 ABSTRACTS Country: Latvia Results: Semen is characterized by low bacteria counts and men without signs of infection may have positive cultures. Occasionally, these microorganisms cause symptomatic urethritis, prostatitis and epididymitis (male accessory gland infection (MAGI)) with deterioration of the semen quality and leucocytospermia. Although CT is a frequent pathogen and a well-documented source of MAGI, the infection does not seem to cause obstruction of the reproductive tract in men, as it does in women. It is postulated that up to 50% of men infected with CT are asymptomatic and in those with symptoms, the most common presentation is urethritis. Discussion: Studies suggest that most upper GTI in young men, also epididymitis, are often attributable to CT. Epididymitis is thought to be important because fertility might be affected due to inflammation, obstruction and functional impairment, especially where both epididymis are affected. As well as creating a blockage to the movement of sperm, CT may also cause epithelial damage that reduces spermatogenesis, induces immunological reactions that hinder sperm, and reduces the female partner's fertility. Conclusions: There are, however, no conclusive studies showing that men infected with CT are less fertile than uninfected men. Male genital chlamydia infection is mainly a threat to the female genital organs. Testing for Chlamydia trachomatis infection in a fertility clinic can be recommended for men in the cases of symptoms of MAGI, leucocytospermia and in the case of positive serological screening for CT in a female partner, if chlamydial infection was not previously diagnosed. CHLAMYDIA ANTIBODIES AS MARKERS FOR LATE COMPLICATIONS IN COST-EFFECTIVENESS MODELS Corresponding author: Jolande A. Land Institution of corresponding author: Division of Reproductive Medicine, Department of Obstetrics and Gynaecology, University Medical Center Groningen, University of Groningen, Groningen, The Netherlands Speciality: Obstetrics and Gynaecology Country: The Netherlands ABSTRACTS E-mail: [email protected] Introduction: Chlamydia infections may give rise to pelvic inflammatory disease (PID) and tubal infertility. The cost-effectiveness of chlamydia screening is determined by the rate of complications prevented. There is a need for more precise estimates of PID and tubal infertility to be inserted in economic models. Chlamydia IgG antibody testing (CAT) has been introduced in fertility clinics as a screening test for tubal infertility. The feasibility of CAT in screening programmes has not been evaluated. Objectives: The aim of our research was to evaluate CAT as an intermediate marker for late sequelae in a cost-effectiveness model. We estimated the prevalence of tubal infertility after chlamydia infection based on data derived from the literature, and compared this with an estimate based on a model including CAT. 40 Methods: We summarized the available literature dealing with the estimates for late complications of chlamydia infections. We constructed a hypothetical model including CAT. For the construction of the model we used data on CAT and tubal pathology in infertile women, and data on seroconversion in PCR-test positive women. Results: In the literature, the generally assumed risk of developing PID after lower genital tract chlamydia infection varies considerably and is up to 30%. The estimated risk for developing tubal infertility after PID is 10-20%. This implies that the risk of PCR-test positive women developing tubal infertility would range between 0.1-6%. In our CAT-based model the estimated a risk of tubal infertility is up to 4.6%. Discussion: The prevalence of CAT positivity is high in infertile women with tubal pathology. Therefore, CAT has been introduced as a screening test in fertility clinics. Based on the results of our study, CAT might be usable in cost-effectiveness studies as an intermediate marker for late sequelae of infection. Research questions that need to be addressed should deal with the pathophysiology of antibody formation in chlamydia infections and the development of tubal scarring. Conclusions: The risk of developing tubal infertility after chlamydia infection appears low (0.1-4.6%). Studies dealing with the transition from lower genital tract infection to tubal infertility are needed to arrive at more precise estimates of the risk of late complications. In these studies, antibody testing as a marker of a previous (asymptomatic) infection alongside PCR testing should be evaluated to conclude whether CAT can be used as a surrogate marker for adverse sequelae in screening programmes. DIAGNOSTICS OF URINOGENITAL CHLAMIDIOSIS COMMON FORMS Corresponding author: Yuriy Vladimirovich Lobzin Other authors: S.N.Sidorchuk, A.I.Poznyak, E.V.Boiko, V.S.Ageev Institution of corresponding author: Medical Academy of Postgraduate Education Institutions of other authors: Military-Medical Academy St.Petersburg Speciality: infectious diseases specialist Country: Russia Introduction: Chronic urogenital chlamydia infection, causes a very wide range of sequelae associated with dissemination of C. trachomatis from the primary focus of infections to sporadic organs. This leads to serious diseases such as genital or extragenital localization (infertility, pregnancy failure, Chlamydia-induced reactive arthritis, oftalmohlamidioz, perihepatitis, pelvic peritonitis). Hematogenous dissemination of agent is the basic unit in the pathogenesis of common forms of chlamydial infection. Objectives: Development of algorithms for diagnosis of chronic chlamydia extragenital localization. 1002 patients at the age of 18-32 were observed and treated for acute urethritis, 41 ABSTRACTS E-mail: [email protected] cervicitis (n=193), localized prostatitis, vesiculoprostatitis, adnexitis, salpingitis (n=340), common Chlamydia trachomatis-induced reactive arthritis (n=274) forms of infection, acute and chronic sinusitis associated with Chlamydia trachomatis (n=95), and meningitis and encephalitis (n=100). Methods: Several techniques were used: polymerase chain reaction (PCR), cell-culture, direct immunofluorescence (DIM), enzyme multiplied immunoassay, indirect immunofluorescence, and electron microscopy. Radioisotope scintigraphy and single-photon emission computed tomography (SPECT) were also carried out. Results: C. trachomatis has been found in the blood of 63.4 % of the patients with confirmed chlamydial infection. Studying the diagnostic importance of detection of the pathogen in blood has shown that in case of chronic genital infections and also extragenital pathologies, the frequency of C. trachomatis detection in serum is 2-3.5 times higher, than in swab material. Discussion: Detection of C.trachomatis in the blood of patients, especially in cases of chronic and complicated forms of chlamydia, has important diagnostic value, because it allows using a fundamentally new approach of the direct detection of the pathogen, regardless of the infection localization. Detection of hlamidemii can be used to diagnose chronic and extragenital forms of chlamydial infection. Conclusions: Using clinical material from the urinogenital tract cannot solve all the diagnostics problems of different urinogenital chlamidiosis forms. Important clinical criterion of common urinogenital chlamidiosis is the presence of chlamydia in blood. To elaborate secondary foci of chlamidiosis it is advisable to use radioisotope scintigraphy. WHAT DO WE REALLY KNOW ABOUT THE SPREAD OF CHLAMYDIA TRACHOMATIS IN COMMUNITIES? Corresponding author: Per-Anders Mårdh Institution of corresponding author: Institute of Clinical Sciences, Department of Obstetrics and Gynecology, Lund, Lund University, Sweden Country: Sweden E-mail: [email protected] ABSTRACTS In many countries, there is no surveillance system for genital chlamydial infections, reason why the extent of carrier ship of Chlamydia trachomatis is unknown. This is then also true for the extent of association of the agent with complications and sequelae that in other studies have been established for Chlamydia trachomatis. How is the situation in countries where genital infections by Chlamydia trachomatis are monitored? The emerging of “the Swedish variant” highlighted that even in countries where such infections are obligatory to report to authorities, the situation is still notably unclear. In surveillance systems, there is generally no distinction between acute and likely chronic infections. There is also no distinction made between the examined number of treated and untreated cases. Likewise, those being tested as notified partners are not reported separately. 42 The importance of understanding the prevalence of genital chlamydial infections in the population to be tested is hardly ever highlighted. In conclusion, a lot more is needed to sophisticate descriptive epidemiological surveys to be able to serve any valuable analytic epidemiological research. COMPETITION OF VACCINE AND NON-VACCINE HUMAN PAPILLOMAVIRUS TYPES BEFORE AND AFTER MASS-VACCINATION Corresponding author: Marko Merikukka Other authors: Johanna Palmroth, Marjo Kaasila, Dan Apter, Jorma Paavonen, Matti Lehtinen Institution of corresponding author: National Institute for Health and Welfare, Oulu, Finland Institutions of other authors: Department of Obstetrics and Gynecology, Kuopio University Hospital, Kuopio, Finland, National Institute for Health and Welfare, Oulu, Finland The Family Federation of Finland, Helsinki, Finland, Department of Obstetrics and Gynecology, University of Helsinki, Helsinki, Finland, School of Public Health, University of Tampere, Tampere, Finland Speciality: Statistics Country: Finland E-mail: [email protected] Introduction: Replacement of multivalent vaccine covered serotypes of pneumococci by non-vaccine serotypes is abolishing effectiveness of pneumococcal mass vaccination. To understand the likelihood of type-replacement following vaccination against human papillomavirus (HPV) types 16/18 we have studied competition of genital HPV types by assessing multiple infections caused by seven most common genital HPV types 6,11,16,18,31,33 and 45 in fertile-aged Finnish females before and after mass-vaccination between 1995 - 2004. Methods: BEFORE mass-vaccination: First trimester serum samples from two consecutive pregnancies (mean 2.5 years apart) were retrieved for a random 3 100 subsample of 123 000 women belonging to the Finnish Maternity Cohort. 42% had antibodies to at least one HPV type at the baseline. AFTER mass-vaccination: In a sizeable phase III (PATRICIA) trial sub-cohort of initially 4808 16-17 year-old Finnish women the HPV16/18 vaccine coverage ranged between <1% and 20% by age-cohort and study community. Results: BEFORE mass-vaccination: Highly significantly increased incidence rate ratios (IRR) of seroconversion to another HPV type were consistently noted for HPV type 33 only, in both HPV16 and HPV18 antibody-positive women: HPV16 antibodies; 16 and 33 antibodies 43 ABSTRACTS Objectives: To explore type-replacement related to HPV mass vaccination using a prophylactic HPV16/18 virus-like particle vaccine (CervarixT) with documented cross-protective efficacy against HPV types 31/ 45, we assessed whether the IRR of non-vaccine HPV types were significantly different in HPV16/ 18 vaccinated women as compared with hepatitis A-vaccine recipients during a 36 month follow-up. (IRR 3.2, 95% CI 2.0-5.2), and HPV18 antibodies; 18 and 33 antibodies (IRR 3.6, 95% CI 2.1-5.9); irrespective of the presence of antibodies to other HPV types at baseline: HPV16 antibodies only; 16 and 33 antibodies (IRR 2.9, 95% CI 1.6-5.4) and HPV18 antibodies only;18 and 33 antibodies (IRR 2.5, 95% CI 1.1-6.0). AFTER mass-vaccination: The IRR estimates of new HPV types acquired during the follow-up in baseline HPV18 positive individuals as compared to baseline HPV18 negative individuals did not differ statistically significantly from 1.0 for non-vaccine, genital HPV types belonging to clades A5, A7, A9 or A10, neither among the HPV16/18 vaccinated nor among the hepatitis A-vaccinated women. Discussion: BEFORE mass-vaccination: A competitive advantage for HPV33 over the other genital HPV types before the era of HPV mass vaccination is suggested. Conclusions: Our studies show no signs of type-replacement following HPV mass vaccination even if some HPV types (HPV33) may have a competitive advantage at the population level in the unvaccinated. THE VALUE OF HOST GENETIC MARKERS COMBINED WITH CT SEROLOGY IN TUBAL PATHOLOGY DIAGNOSIS Corresponding authors: Sander Ouburg, Servaas A. Morré and the FP6 EpiGenChlamydia Consortium Institution of corresponding authors: Laboratory of Immunogenetics, Department of Pathology, VU University Medical Center, Amsterdam, The Netherlands Country: Netherlands e-mail: [email protected] ABSTRACTS Chlamydia trachomatis (CT) infections are the world leading cause of preventable blindness (trachoma) and the most prevalent sexually transmitted disease of bacterial origin which is strongly associated with pelvic inflammatory disease, ectopic pregnancy, and tubal infertility. Twin study-based findings of members of the FP6 EpiGenChlamydia Consortium identified a 40% genetic predisposition to CT infections. The EpiGenChlamydia Consortium structured transnational research to such degree that comparative genomics and genetic epidemiology on large numbers of unrelated individuals can be performed to identify genetic markers to be used for patient profiling. Subfertility poses an enormous burden on healthcare and society throughout the world. Worldwide, 15% of couples trying to conceive suffer from subfertility. Subfertility generally described as a failure to conceive after one year of unprotected, regular sexual intercourse. Approximately half of the couples suffering from subfertility will conceive spontaneously, or after simple treatment. The other half needs more complex treatment, such as in vitro fertilization (IVF) or other assisted reproduction procedures. There are several causes of subfertility, which can be classified as ovulation disorders, male factor subfertility, tubal damage, unexplained subfertility, and other causes, such as endometriosis and fibroids. In women, one of the major causes of subfertility is tubal pathology, with a prevalence of around 30%. In all of cases of tubal pathology, C. trachomatis is the most common cause for infertility. 44 The reference standard for diagnosing tubal pathology in subfertile women is laparoscopy performed usually after a CT IgG positive antibody test which is only positive in 50-60% of women with tubal pathology. Despite the effectiveness, laparoscopy is costly, invasive and has a low positive predictive value. Therefore there is a unmet medical need for better diagnosing tubal pathology in women with subfertility, a need which could potentially be met by adding genetic profiling to current diagnostic approaches. The current presentation will discuss immunopathological mechanisms and host based genetic diagnostic approaches. PREVENTION OF CT AND HPV INFECTIONS: ARE WE DOING ENOUGH? Corresponding author: Jorma Paavonen Institution of corresponding author: Department of Obstetrics and Gynecology, University Hospital, Helsinki, Finland Country: Finland E-mail: [email protected] Chlamydia trachomatis (CT) infections of the genital tract are highly prevalent globally. CT is a true obligate intracellular pathogen which has a unique growth cycle distinguished from all other micro-organisms. Because of this slow growth cycle chlamydial infections tend to be silent and chronic. The clinical spectrum of CT infections is wide. Most clinical manifestations of CT genital infections are asymptomatic. Secondary prevention by screening for chlamydial infections still remains important. CT vaccine development has proven difficult because of the complex antigenic structure and limited knowledge of the protective antigens. Guidelines or recommendations for annual screening of young sexually active individuals for chlamydial genital infections have been implemented in many countries. However, the effectiveness of screening programs has recently been challenged since population studies suggest that screening programs have been strikingly ineffective in lowering chlamydia rates. By contrast, the rates of reported chlamydial infections have been increasing in many countries regardless of implementation of screening. Human papillomavirus (HPV) infection is the most common viral sexually transmitted infection. HPV infections are extremely common, and most young adults are exposed to HPV soon after sexual debut. Although most HPV infections resolve, persistent infection by one or more of the oncogenic HPV types can cause cervical neoplasia. HPV can also cause other anogenital neoplasias and a smaller proportion of oropharyngeal neoplasias. Worldwide, cervical cancer is the second most common cancer in women and a major cause of cancer death in women worldwide. Secondary prevention by cytologic screening has been effective in some countries, but the screening policies vary widely. Therefore, the incidence rates of cervical cancer differ strikingly between countries, also within Europe. Due to mass screening, the disease burden has shifted to management of cervical intraepithelial 45 ABSTRACTS Thus, there is major frustration with opportunistic or organised screening programs, implementation of management guidelines and contact tracing efforts. Screening activity may certainly benefit individuals but seems to have little public health impact. neoplasias (CIN). This causes health problems and drains health care resources. The development of prophylactic HPV vaccines has been a remarkable success story. Primary prevention by these vaccines can substantially reduce the public health and economic burden of cervical precancer and cancer and other HPV-associated diseases. Clinical efficacy trials have demonstrated that these vaccines are strikingly effective in the prevention of high grade cervical, vaginal and vulvar precancers. These trials have also raised several questions and challenges to be addressed to assess the public health impact on all HPV-associated diseases, and to design the most effective vaccination strategy. These include duration of immune response, vaccination of males, efficacy against other HPV-related cancers bewyond cervical cancer, postmarketing safety surveillance, impact of vaccination on existing screening programs, potential HPV type-replacement after widespread vaccination, and finally feasibility of HPV vaccination in developing countries where the disease burden remains enormous. Although major improvement has taken place in recent years in our armamentarium to prevent sexually transmitted CT and HPV infections the question still remains: Are we doing enough? EPIDEMIOLOGICAL SURVEILLANCE OF CHLAMYDIA TRACHOMATIS INFECTION IN LATVIA Corresponding author: Jurijs Perevoscikovs Other authors: Violeta Mavcutko Institution of corresponding author: Epidemiological Safety and Public Health Department, State Agency "Infectology Center of Latvia" Institutions of other authors: HIV/ AIDS Surveillance and Prevention Unit, Epidemiological Safety and Public Health Department, State Agency "Infectology Center of Latvia" Speciality: epidemiology Country: Latvia ABSTRACTS E-mail: [email protected] Introduction: Chlamydia trachomatis is the most commonly reported sexually transmitted infection (STI) in Latvia. The proportion of Chlamydia infection increased in STI structure up to 66% last year. Genital Chlamydia infection is a significant public health concern because untreated infections cause complications with serious consequences on women's reproductive health. There are four STI under mandatory notification in Latvia. STI are in the 4th place in the structure of the notifiable infections in Latvia. Objectives: Epidemiological surveillance system monitors the spread of Chlamydia trachomatis and provides important information for effective control of the infection. Chlamydia is easily treated with antibiotics and a preventable disease (safe sex, condom use). An important aspect of prevention involves the evaluation of sexual partners to prevent re-infection and further spread of the disease. 46 Methods: C. trachomatis is a mandatory notifiable infection in Latvia. Data from the national computer-based surveillance system VISUMS were used to describe epidemiological trends for the period from 2008 to 2010. The case definitions used in Latvia are based on the European Union case definitions. The reports of Chlamydia cases to the national surveillance system contain individual laboratory notification from a diagnostic laboratory and an individual clinical notification from health care providers. Results: In Latvia, altogether 2873 Chlamydia cases (58.5% of all cases of STI) were reported to the national surveillance system from 2008 to 2010. The proportion of Chlamydia in STI structure increased from 46% in 2008 to 66% in 2010. Up to 66% of all cases were reported in the age group 18 - 29 years. The proportion of females in structure of Chlamydia infection increased from 46% in 2008 to 64% in 2010. In women of age group 18-29 years the incidence rate of genital Chlamydia increased by 2,2 times, from 105,1/100 000 in 2008 to 238,4/100 000 in 2010. During the period of 2009 to 2010 reported cases in pregnant women increased from 57 (9.4% of all reported females cases) to 174 (26%) cases. Discussion: In Latvia the highest incidence rate of Chlamydia infection was found in young women of reproductive age. Chlamydia is usually asymptomatic in both women and men. In women, it may result with complications, which is a major cause of tubal infertility, ectopic pregnancy. The number of diagnosed and treated cases depends on the number of persons tested. In Latvia legislation obliges testing for asymptomatic cases only for pregnant women, but there is a lack of screening program for populations. Conclusions: During the last two years the number of notified Chlamydia cases in females notably increased (by 206%). It could be explained by the following reasons: since July 2008 all microbiology laboratories were involved in notification system and significant effort has been made to improve the completeness of reporting by health care providers in Latvia. The development and implementation of a screening programme is very important for further improvement of surveillance and control of Chlamydia infection. GENOTYPING OF CHLAMYDIA TRACHOMATIS STRAINS Corresponding author: Mirja Puolakkainen Other authors: Suvi Niemi and Eija Hiltunen-Back Institutions of other authors: Haartman Institute, Department of Virology, University of Helsinki, and HUSLAB, Department of Virology and Immunology, Helsinki University Central Hospital, Clinic of Venereal Diseases, Skin and Allergy Hospital, Helsinki University Central Hospital, Finland; National Institute of Health and Welfare, Helsinki, Finland; Helsinki, Finland Speciality: clinical microbiology Country: Finland E-mail: [email protected] 47 ABSTRACTS Institution of corresponding author: Haartman Institute, Department of Virology, University of Helsinki, and HUSLAB, Department of Virology and Immunology, Helsinki University Central Hospital, Helsinki, Finland Introduction: Since 1995, the number of Chlamydia trachomatis infections has been officially notified in Finland. Lately, there has been around 14 000 notified cases annually (250 cases/ 100 000 inhabitants). C. trachomatis infections are diagnosed with sensitive and specific nucleic acid amplification tests (NAATs), but these tests do not differentiate between genotypes. We wanted to further refine the current laboratory diagnosis of C. trachomatis infections. Objectives: Our purpose was to study molecular epidemiology of urogenital and extragenital C. trachomatis strains, and to study the occurrence of the Swedish new variant of C. trachomatis in Finland. Methods: We set up real-time quantitative PCR based methods for genotyping C. trachomatis types D-K (ompA-PCR) and L1-L3 (pmpH-PCR) as well as the Swedish plasmid variant of C. trachomatis (plasmid-PCR). Results: Of the 160 C. trachomatis positive urogenital samples analyzed, 90% could be genotyped with the ompA real-time PCR. The three most prevalent genotypes were E, F and G. No L types were detected. Although the genotype E was the most common genotype among C. trachomatis strains, the Swedish new variant was rarely found (only 2/469). Of the 100 C. trachomatis strains detected in extragenital specimens, 79 could be genotyped. Five rectal swab specimens harbored the genotype L2b. All these patients belonged to the identified risk groups. Discussion: Typing of C. trachomatis strains can be used in epidemiologic surveillance, to assess the changes in genotype distribution and to reveal transmission patterns in sexual networks. This information is of value in efforts to monitor and prevent spread of C. trachomatis infections. Conclusions: The most prevalent genotype in Finland was E (40%). The Swedish variant was rare in Finland despite our close proximity to Sweden. Also in Finland, L2b genotypes were detected in rectal swabs taken from HIV positive MSM. NEW METHODS FOR CERVICAL CANCER DETECTION IN LITHUANIA. VALUE OF HIGH-RISK HPV-DNA TESTING IN THE TRIAGE OF WOMEN WITH ASC CATEGORY FROM LIQUID BASED CYTOLOGY Corresponding author: Jolita Rimiene Institution of corresponding author: Diagnostics Pathology, Vilnius University Country: Lithuania ABSTRACTS E-mail: [email protected] Background: Cervical cancer in Lithuania is the fourth commonest cancer among women with malignancies. Since 1992 Lithuania have the increasing rates in incidence of cervical cancer among Europe countries and the highest in Baltic sea region countries. The Lithuanian ministry of Health started nation- wide cervical cancer screening program in the middle of 2004. During the program overall was diagnosed 7 778 HSIL and 323 cancer cases. In the screening group during program period we observed statistically significant increase in incidence of stage I cervical cancer and decrease in stage II. The changes in incidence of stages III and IV cervical carcinoma we did not observed. During the program period we observed statistically significant increased incidence of CIN3/CIS – from 1,7/100 000 in 1999 to 31,1/100 000 in 2008. The incidence was increasing by 34,4% in all age group and by 48 33,7% in 25–59 age group and by 26,4% in 60 and older years age group. Cytological screening has reduced the incidence of invasive cervical cancer, but as a result of screening, many women are diagnosed as having equivocal cytological abnormalities (eg, atypical squamous cells, herein referred to as ASC). 11 pathology laboratories are accredited to perform conventional Pap tests. According Screening Program period in Lithuania was diagnosed 16 545 (3%) ASC cases, high number of unsatisfactory smears (2,3-9,6%). Many laboratories are performing HPV testing from fresh cytological material after the conventional Pap smear. We introduce Cobas® 480 HPV detection system in combination with ThinPrepTM liquid based cytology. We analyzed the initial data from the first 1397 ThinPrepTM Pap tests. 13.7% abnormal cytology cases were found. Low 0.6% unsatisfactory and 1,1% ASC ratio was observed. A high number of LSIL and HSIL (8% and 4.6%) cases was detected. Endocervical adenocarcinoma case was diagnosed. The difference number of SIL cases was detected in healthy women group and women with risc factors: 5.7% LSIL and 2.5% HSIL, 7% LSIL and 7% HSIL respectively. HPV infection was found in 39 (73.6%) of 53 tested HSIL and 14 (77.8%) of 18 LSIL ThinPrepTM vials. HPV was detected in 8 (47%) of 17 ASC cases. In 5 ASC-H and 3 ASC-US cases HR-HPV types was detected. 4 of 5 women who underwent conisation CIN2/CIN3 lesion was found in the histological material. Conclusions. Liquid based cytology and reflex HPV DNA testing is effective measure for early cervical cancer detection. HPV-DNA testing may serve as a means to better select which patients with ASC-H and ASC-US on Pap cytology should undergo colposcopic examination and conisation. This approach potentially may reduce medical costs and eliminate the need for routine repeat cytology, colposcopic examination among patients with ASC-US Pap smears. DO ABBOTT REALTIME CT AND -HIGH RISK HPV MEET THE REQUIREMENTS FOR LABORATORY SCREENING AND DIAGNOSIS OF CHLAMYDIA TRACHOMATIS AND HUMAN PAPILLOMA VIRUSES? Corresponding author: Edwin Roovers Institution of corresponding author: Abbott Molecular Speciality: Marketing Manager EMEA&India Country: The Netherlands Introduction: Various forms of Chlamydia screening using NAAT have been suggested but they were implemented only in a limited number of countries. HPV DNA testing has become an important part of cervical carcinoma screening and management algorithms in several countries. Clinical trials were conducted to show the performance of the RealTime CT and RealTime High Risk HPV assays as well as how they compare versus other methods. Also more practical criteria for the laboratory will be discussed. Methods: The m2000 system is a fully automated IVD system for nucleic acid extraction, real-time PCR and results reporting. A multicenter clinical study was conducted to evaluate the performance characteristics of the Abbott Real-Time CT/NG assay compared to Aptima Combo 2 Assay (Gen-Probe), ProbeTec ET CT/GC (BD). For HPV the clinical performance of the Abbott Real-Time High Risk HPV test and Hybrid Capture 2 HPV DNA Test was prospectively compared in a screening setting. 49 ABSTRACTS E-mail: [email protected] Conclusions: The Abbott m2000 provides laboratories with ease of use, reliability and efficiency for a broad spectrum of IVD assays. It is also an open system for running in house methods. Taking factors into consideration such as assay sensitivity & specificity, accuracy, efficiency and reliability it can be confirmed that Real-Time CT and Real-Time High Risk HPV do meet the high standard requirements for laboratory screening and diagnosis of Chlamydia trachomatis and human papillomaviruses. IL6 -174 G>C POLYMORPHISM, HPV INFECTION AND CERVICAL INTRAEPITHELIAL NEOPLASIA Corresponding author: Iona Rotar1 Other authors: Daniel MURESAN1, Radu POPP2, Sorana D. BOLBOACA3, Felicia PETRISOR2, Cristina BUTUZA4, Silvana APOSTOL4, Georgiana COROIU5, Florin V. STAMATIAN1 Institutions of other authors: 1- „Iuliu Hatieganu” University of Medicine and Pharmacy Cluj-Napoca, 1st Department of Obstetrics and Gynecology, 3-5 Clinicilor, 400006 Cluj-Napoca, Romania Country: Romania E-mail: [email protected] Introduction: The relationship between chronic inflammation and cancer is well-know especially for HPV and cervical cancer. Interleukin-6 (IL-6) an inflammatory cytokine can plays either inhibiting or stimulating roles in carcinogenesis. Objective: In this study we aim to analyze the relation between cervical cancer and IL6 -174 G>C polymorphism. Materials and methods: A case control study has been conceived. The cases were represented by 128 patients diagnosed with cervical intraepithelial neoplasia, positive at HPV HR testing, while 111 controls, negative for intraepithelial neoplasia and negative at HPV HR testing were included in the control group. For each patient a physical examination has been performed. Both cervical and peripheral blood sample were obtained. From the cervical probe cytology and HPV HR testing has been done, while DNA has been extracted from blood. Later IL6 -174 genotyping was detected using a PCR-RFLP technique. ABSTRACTS Results: The absolute genotypes frequencies were for cases: G/G - 60, C/G – 52, C/C- 16 respectively for controls G/G – 19, G/C- 47, C/C – 19. Chi square had a value of χ2 = 0.968; df = 1; p = 0.325 while considering G/C&/C/C genotype as a risk factor, respectively χ2 = 1.014; df = 1; p = 0.314 when only homozygous C/C genotype was taking into consideration as a risk factor. OR had values of 1.2, CI 95% [0.774, 2.163] for G/C&/C/C genotype, respectively OR-1.446, CI 95% [0.704, 2.970] for T/T genotype considered as a risk factor. Discussion: HPV represents a mandatory condition for cervical carcinogenesis. In particular the genetic polymorphism at -174 could be followed by a change in IL-6 local and serum level. By the binding of the latter interleukin to its receptor a Jak pathway is activated. The implication of Jak kinase has been previously reported especially for hematologic cancer but also for solid carcinomas. Conclusion: The presence of at least one variant allele could be considered as a risk factor for cervical intraepithelial neoplasia but the association is weak. 50 WHAT SHOULD BE THE BASIS OF A NEW CERVICAL SCREENING PROGRAMME? Corresponding author: Peter Sasieni Country: United Kingdom E-mail: [email protected] Cervical cancer rates have been greatly reduced in many industrialized countries through screening programmes based on cervical cytology. Unfortunately this technology has had little impact in low and middle income countries. Today HPV testing and visual inspection methods are available as alternatives to cytology. Additionally, HPV vaccination offers a one-off approach to cervical cancer prevention. I will consider the pros and cons of different approaches to screening and argue that except in places with an established and effective cytology based screening programme, screening should be based on HPV testing with other tests reserved for triage of HPV positive women. Whilst vaccination may be seen as the solution for the future, a need for good screening will remain for many years to come. HPV SURVEILLANCE IN THE VACCINE ERA Corresponding author: EJ Savage Other authors: R Howell-Jones, K Soldan Institution of corresponding author: Health Protection Agency Specialty: STI surveillance Country: United Kingdom Introduction: An important component of any vaccination programme is monitoring to assess the impact of the programme. The Human papillomavirus (HPV) vaccine was introduced into the UK immunisation programme in 2008 and is offered to all females aged 12-13 years of age and a “catch-up” campaign was conducted to reach those aged 14 to 18 years. High levels of uptake have been achieved with 76.4% of 12-13 year old females completing the three dose course in 2009/10. Cervarix, the vaccine currently used in the UK programme, protects against HPV types 16 and 18 which are associated with over 70% of cervical cancers in the UK. Conclusions: We will present an overview of HPV surveillance programmes that have been established to monitor key outcomes with a particular focus on England. As well as vaccine safety and coverage, systems are now in place to monitor changes in the frequency of vaccine (and non-vaccine) HPV types, vaccine effectiveness, immunogenicity and herd-immunity. 51 ABSTRACTS Email: [email protected] THE AMAZING STORY OF THE SWEDISH NEW VARIANT OF CHLAMYDIA TRACHOMATIS (nvCT) Corresponding author: Magnus Unemo Institution of corresponding author: National Reference Laboratory for Pathogenic Neisseria, Department of Laboratory Medicine, Microbiology, Örebro University Hospital, Örebro, Sweden Speciality: Microbiology and Molecular Biology Country: Sweden Email: [email protected] Introduction: In late-2006, the new variant of Chlamydia trachomatis (nvCT) was reported in Sweden. The nvCT contains a 377 bp deletion within its plasmid, which encompasses the targets in the commercial diagnostic PCR systems from Roche and Abbott at the time. The nvCT rapidly spread nationwide in Sweden, undiagnosed by these systems, and caused thousands of false negative reports. Objective: To provide a comprehensive overview of all published data regarding nvCT, its emergence, epidemiology, phenotypic and genetic characteristics, and effects on public health and C. trachomatis surveillance. ABSTRACTS Overview: The nvCT is clonal and has been assigned serovar/genovar E, multilocus sequence typing [MLST] ST 21 (hctB), 19 (CT058), 1 (CT144), 2 (CT172), and 1 (pbpB), as well as variable number of tandem repeats [VNTR] type 8.7.1 (CT1335, CT1299, and CT1291, respectively). Genome sequencing and detailed phenotypic characterisation have shown that the biological fitness of nvCT in vitro, when compared to wild type CT strains, is unaltered. This is supported by epidemiological and most clinical observations. Accordingly, the initial rapid transmission of nvCT in Sweden was due to the diagnostic selective advantage and its introduction into a high-frequency transmitting population. The proportions of nvCT cases now appear to be converging towards equilibrium with the wild type CT strains in the different Swedish counties. The absence of any isolates from before 2000 and statistical estimations suggest that nvCT emerged 4–6 years before it was detected. Interestingly, the nvCT remains rarely reported beyond the Nordic countries and the epidemiological reasons for this finding are not elucidated. Summary: The emergence of nvCT in Sweden created a unique opportunity to study the emergence, spread and effects on public health of a single strain within both the human and bacterial populations. The nvCT had a substantial impact on C. trachomatis identification, epidemiology, and public health in Sweden. The lessons learned, which are also applicable to other infectious diseases, include; importance of epidemiological surveillance and especially detailed investigation of changes in the incidence and epidemiology of infection, the frequent participation in appropriate quality assurance schemes and the careful design of diagnostic assays (ideally with multiple targets). It is crucial to have diverse diagnostic systems and response plans prepared for similar situations in the future. 52 FIRST SCREENING PROJECT REGARDING CHLAMYDIA TRACHOMATIS INFECTION IN ALBANIA Corresponding author: Adela Vasili Other authors: Lila SHUNDI, Brunilda VILA, Silva BINO, Eugena ERINDI, Gjergji KOJA, Dritan ULQINAKU, Shpetim QYRA, Mimoza BASHO Institution of corresponding author: Institute of Public Health, Tirana, Albania Institutions of other authors: Institute of Public Health, Tirana, Albania Speciality: Epidemiologist Country: Albania E-mail: [email protected] Introduction: Chlamydia trachomatis infections are now recognized as the second most frequent cause of sexually transmitted diseases (STD) worldwide. Considering that in Albania, till now, a national control program on genital chlamydial infection has not been implemented, the development of a screening project in Albania was necessary. Objective: The aim of this study was to determine the prevalence of Chlamydia trachomatis genital infections, as well as the feasibility of molecular screening in Albanian women of reproductive age. Materials and Method. Study population: From October to December 2010, endocervical swab specimens were collected from 241 sexually active women, under 40 years and consulting in women’s health services in 4 districts Tirana, Durres, Vlore and Fier. The swabs were collected using Amplicor STD Swab Specimen Collection and Transport Kit (Roche Diagnostics Systems). The selected women filled in a standardized questionnaire. Molecular amplification analysis was performed on all samples using Cobas AmplicorTM CT Test (Roche Diagnostics Systems) on the COBAS Amplicor instrument, following the manufacturer’s instructions. Conclusion: This study showed the prevalence and revealed the risk factors related to C. trachomatis infections in Albania. This study, being the first of this kind in Albania, used a nucleic acid amplification test for the identification C. trachomatis in a selected population. This study creates the opportunity for implementing a national control and prevention program on genital Chlamydial infection, which does not exist so far. Additional information: In Albania there is no data on the incidence of chlamydial infection, which is not mandatorily to be reported by the laboratory or clinic. In Albania there was a national program to control and prevent HIV-AIDS, but for chlamydial infections, this is the first initiative. 53 ABSTRACTS Results: A total of 241 women of median age 30 years (range 17-40 years) were studied. The prevalence rate was 7.5%, (95% CI=4.5%-11.5%). The prevalence differed significantly by age: less than 20 years the prevalence is 0; 11.9% (5/42), among women 20-24 years, 9.1% (6/66) for the women 25-29 years and 5.8% (7/121) among those over 30 years. A new sexual partner in the last 6 months was the main risk factor to be C. trachomatis infected with an OR 7.2 (95% CI=2.38-23.2). Neither reason for consulting nor the existences of symptoms were significantly associated with being C. trachomatis infected. WWW.GEN-PROBE.COM GP-Ads.indd 1 4/1/11 4:38 PM WWW.GEN-PROBE.COM GP-Ads.indd 2 4/1/11 4:38 PM RAPID DIAGNOSTICS FOR CHLAMYDIA TRACHOMATIS Corresponding author: Abhay Vats Other authors: Ashok Gurung, Ashish Yeri, Di Gao, David Mead, Toni Darville, Shiela West, Ishwad Chandra Institution of corresponding author: University of Pittsburgh Institutions of other authors: Johns Hopkins University, Lucigen Corp Speciality: Pediatrics Country: USA E-mail: [email protected] Introduction: Chlamydia infections are a major cause of morbidity and preventable blindness in the developing world. In addition, they represent a major cause of sexually transmitted diseases worldwide, and many such cases can be difficult to diagnose due to lack of appropriate testing facilities in poor resource settings. Objectives: We aimed to develop a low cost detection method for Chlamydia trachomatis, which would be useful for both eye and genital infections. It is based on Loop mediated isothermal DNA amplification technique (LAMP). Methods: We developed primers and amplification protocols for several targets in the Chlamydia genome as well as a lateral flow detection method for amplicons. Primers were labeled with FITC, Biotin and Digoxigenin for detection in the lateral flow strips. A self contained cassette was used for detection of the LAMP amplicons. Results: The method was able to successfully amplify Chlamydia DNA using a low cost heater (heater cost <$2.00). The amplicons were detectable within 5 minutes after the reaction on the lateral flow strips. The whole procedure takes less than 30 minutes and cost less than $5.00 per reaction with sensitivity and specificity similar to PCR. Discussion: We have developed a low cost and highly sensitive and specific amplification and detection system for Chlamydia trachomatis diagnostics. ABSTRACTS Conclusions: Isothermal DNA amplification and lateral flow detection methods could be useful for managing this infection in low resource settings. Further development and clinical testing of the assay is being planned. 56 EPIDEMIOLOGY AND CONTROL OF GENITAL CHLAMYDIA TRACHOMATIS INFECTION IN SWEDEN Corresponding author: Inga Velicko1,2 Other authors: Kühlmann-Berenzon S1 Institution of corresponding author: 1- Department of Analysis and Prevention, Swedish Institute for Communicable Disease Control, Solna, Sweden 2- Department of Medical Epidemiology and Biostatistics, Karolinska Institutet, Stockholm, Sweden Institutions of other authors: 1- Department of Analysis and Prevention, Swedish Institute for Communicable Disease Control, Solna, Sweden Speciality: Epidemiologist Country: Sweden E-mail: [email protected] ABSTRACTS Genital Chlamydia infection is mandatory notifiable in Sweden through the coded, anonymous electronic web-based notification. The majority (86%) of all reported cases is between the ages of 15 and 29 years, and 57% are females. Chlamydia incidence increased significantly between 1997 and 2004, from 157 to 356 cases per 100 000 inhabitants. The following emergence of a new genetic variant of Chlamydia trachomatis in 2006, distorted the “natural” history of Chlamydia epidemiology as cases were missed due to the inability of some of the diagnostic-kits to detect this new variant. The catch-up of cases that followed imitated an artificial increase in incidence that was clearly observed in 2007 and 2008. Chlamydia incidence is significantly dependant on testing activity, which has increased in Sweden continuously since 1997. In 2009 more than half a million persons were tested, and 7% of those were positive. However, only one third of tested persons were males. Given the figures and the limited testing coverage, it is safe to say that Sweden does not yet have an effective control on the spread of the disease. For this reason, and based on the epidemiologic background and data from past surveys on sexual health, the authorities initiated a “National Programme for chlamydia prevention with focus on adolescents and young adults, 2009-2014”. The programme’s measures are expected to show an effect after some years and should therefore be critically and continuously evaluated in order to incorporate adjustments as needed. 57 Molecular diagnostics Finding answers, innovating solutions STDs 4000 INFANTS BLINDED ANNUALLY Innovating medical devices and molecular diagnostic solutions ABSTRACTS How some untreated and undetected STDs contribute to blindness in more than 4,000 babies every year. It is tragic. And Needless. BD offers a highspeed molecular-based solutions as an answer to an increasing demand for testing of CT/GC, HSV 1&2 and HPV* at a time when labs are challenged by decreased healthcare costs enforcing them to deliver more and higher quality results with fewer resources. For more information, please visit: www.bd.com * products In development 58 BD and BD Logo are trademarks of Becton, Dickinson and Company. ©2011 BD Printed in Europe. Becton Dickinson GmbH Tullastrasse 8-12, D-69126 Heidelberg. Germany. Sitz: Heidelberg. Amtsgericht Mannheim HRB 330 707. flier.indd 1 08/04/2011 15:01:25 SELF-SAMPLE HPV-TESTS - EFFECTS IN CERVICAL CANCER SCREENING ATTENDANCE AND COVERAGE IN FINLAND Corresponding author: Anni Virtanen Other authors: Pekka Nieminen, Tapio Luostarinen, Ahti Anttila Institution of corresponding author: Mass Screening Registry, Finnish Cancer Registry Institutions of other authors: Department of Obstetrics and Gynaecology, Helsinki University Central Hospital, Jorvi Hospital Mass Screening Registry, Finnish Cancer Registry Speciality: cervical cancer screening Country: Finland E-mail: [email protected] Introduction: Attendance in screening is an important determinant of cervical cancer. In Finland, the overall attendance rate in the organized screening programme is 70% and attendance among women aged 30-35 only 50-60%. High-risk human papillomavirus (hrHPV) DNA detection based testing on patient obtained samples (self-sampling tests) are sensitive in finding cervical cancer and its precursors. Previous experience also suggests a good effect among non-attendees of screening. Objectives: We assessed the effects of self-sampling among the non-attendees of the routine screening programme of a Finnish municipality. Main outcomes were increases in screening attendance and coverage (coverage of any Pap-smear within the 5-year screening interval) achieved by self-sampling kits, reminder letters or the combination of these two. Additional outcome was to compare the prevalence of hrHPV test-positivity among first invitation and second/third intervention attendees. Methods: Non-attendees after the primary invitation were randomised to receive either a self-sampling kit (2,397 women) or a reminder letter (6,302 women). One fourth (1,320 women) of arm non-attendees after a reminder letter also received a self-sampling kit as a third intervention. Effects on screening coverage were assessed according to the self-reported previous Pap smear history of the participants. Discussion: Our results suggest that if self-sampling serves as the third intervention after two reminder letters, on a national scale an increase in participation rate of organized screening is achievable from 70 to 80% or more. Self-obtained samples are most likely more often HPV-positive because they also reveal vaginal or even vulvar HPV infections in addition to the cervical ones. Conclusions: In increasing the attendance at organized screening, self-sampling can feasibly be used as an option for a reminder letter, or as an addition to it. More research is needed to assess effects in different settings, as well as with regard to prevented cancers and cost-effectiveness. 59 ABSTRACTS Results: Participation rate by self-sampling, 31.5% (CI: 29.7, 33.4%), was significantly higher than with a reminder letter, 25.9% (CI: 24.8, 26.9%). The adjusted relative risk for participation by self-sampling was 1.21 (CI 1.13-1.3). Total attendance increased from 64.9 to 76.0% with self-sampling and from 64.6 to 73.8% with a reminder letter. Combining the interventions reached 40% of non-attendees and increased total attendance from 63% to 78%. Only app. 20% of the participants in all three groups increased screening coverage (previous Pap-smear ≥5 years ago or never). Self-obtained samples were more often HPV-positive than provider obtained ones, 12-13% versus 7-8%. Abbott RealTime High Risk HPV • One Assay – Three Results: – Detection of 14 HPV high-risk types – Genotyping of HPV 16 – Genotyping of HPV 18 • More confidence in detection of disease compared to cytology* • HPV-typing results will help guide clinical management decisions** Flexible Sample Collection Options for RealTime High Risk HPV on m2000 system • Validated with ThinPrep PreservCyt Solution • Validated with SurePath Preservative Fluid (pre/post-gradient) AM0019EU2011 • Validated with Abbott Cervi-Collect – simplifying laboratory workflow 310473_Eurogin_Anzeige.indd 1 * Huang et al. Journal of Clinical Virology, 2009; 45 (Suppl 1): 19 – 23 ** Kahn et al. Journal of the National Cancer Institute, 2005; 97 (14):1072 – 1079 17.03.11 14:18 Addressing Chlamydia testing needs Abbott RealTime CT & CT/NG Simplicity One ready-to-use collection device for multiple specimen types Sample Validity High extraction quality with true internal control Flexibility on One Platform Broad IVD assay menu and laboratory defined assays Walk-Away Automation Low to high throughput options We Listen. www.abbottmolecular.com Abbott RealTime is a trademark of Abbott Laboratories in various jurisdictions. Ad_CTNG_2009_V5.indd 1 10.05.2009 19:14:20 MAIN SPONSOR PLATINIUM SPONSOR SPONSORS Conference webpage: www.cthpv.org
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