Anti-Chlamydia trachomatis IIFT Positive reaction Negative reaction Antibodies against Chlamydia trachomatis Indications: Test system for the in vitro determination of antibodies against Chlamydia trachomatis in human serum or plasma for the diagnosis of the following diseases: trachoma, urogenital tract infections and lymphogranuloma venereum. Clinical significance: The infectious agent Chlamydia trachomatis belongs to the human pathogenic Chlamydia genus, together with Chlamydia pneumoniae and Chlamydia psittaci. It is one of the smallest intracellular gram-negative bacteria. Around 700 million people are infected worldwide, with an annual new infection rate of around 50 million. The disease is transmitted by contact with infected humans. C. trachomatis is the agent of non-gonorrheal urethritis, lymphogranuloma venereum, trachoma, inclusion conjunctivitis, neonatal pneumonia and Reiter’s syndrome. Sexually transmitted non-gonorrheal urethritis is the most frequent venereal disease. In Germany the C. trachomatis serotypes D to K are responsible. They cause urethritis, epididymitis and prostatitis in men, and urethritis, cervicitis and salpingitis/adnexitis in women. Lymphogranuloma venereum is caused by the Chlamydia trachomatis serotypes L1 to L3. It is a rare venereal disease which occurs worldwide but mainly in tropical areas. In tropical regions, C. trachomatis leads to trachoma (with serotypes A, B, Ba and C), an eye infection of varying severity. It is caused by direct contact with the mucous membranes of the eye, nose and mouth and can be transmitted by common use of hand towels. Around 400 million people suffer from trachoma, which is worldwide the most frequent cause of blindness (trachoma blindness). In newborns, particularly premature infants, prenatally or perinatally transmitted C. trachomatis causes pneumonia (serotypes D to K) in addition to conjunctivitis (ophthalmia neonatorum). This is noticeably often accompanied by pneumothorax and lifelong health problems. Despite distinct clinical symptoms, C. trachomatis antibodies are not necessarily produced in the serum in localised processes. IgM antibodies are not formed in all cases of florid infection. An increase in IgG titer is also not always found. In problem cases, it is therefore useful to determine the presence of Chlamydia in infectious secretions using direct immunofluorescence or to determine their specific genetic sequences using PCR, which is often unsuccessful in cases where the infection has taken place some time ago. The fastest and most reliable investigation of specific antibodies in infections with C. trachomatis, even with low antibody titers, is obtained with MIF (microimmunofluorescence) as “gold standard“, IIFT (indirect immunofluorescence test) and ELISA (enzyme linked immunosorbent assay) using MOMP as the target antigen and taking into consideration different serotypes. Studies throughout Europe show that these methods are suited to confirming C. trachomatis-induced infertility in women and men by the determination of C. trachomatis specific serum IgA and IgG antibodies. Recognised medical centres therefore recommend C. trachomatis screening for both parents before pregnancy. Fluorescence pattern (positive reaction): Antibodies against Chlamydia trachomatis cause a fluorescence of the inclusion bodies located in the cytoplasm. These inclusion particles contain densely organised elementary bodies (infectious form of Chlamydia, diameter 300 nm) and reticular bodies (non-infectious form of Chlamydia, diameter 1000 nm). Free elementary and reticular bodies lying between the cells can also react. Some of the cells in the field of view are not infected and show no fluorescence. EUROIMMUN IIFT Infectious Serology Viruses: Adenoviruses Chikungunya virus Coxsackieviruses Crimean Congo fever virus (CCHFV)* Cytomegalovirus (CMV) Dengue viruses types 1-4 (DENV) ECHO virus Epstein-Barr virus capsid antigen (EBV-CA) Epstein-Barr virus early antigen (EBV.EA) Epstein-Barr virus nuclear antigen (EBNA) Hantaviruses (types Hantaan, Puumala, Seou Saaremaa, Dobrava, Sin Nombre, Andes)* Herpes simplex virus types 1 and 2 (HSV-1/2) Human herpes virus type 6 (HHV-6) HIV-1 and -2* Influenza virus types A and B Japanese encephalitis virus (JEV)* Measles virus Mumps virus Parainfluenza viruses types 1-4 Respiratory syncytial virus (RSV) Rubella virus* Sandfly fever virus* (types Sicilian, Naples, Toscana, Cyprus) SARS Coronavirus (SARS-CoV) Tick-borne encephalitis (TBE) virus Varicella zoster virus (VZV) West Nile virus (WNV) Yellow fever virus (YFV) Bacteria: Afipia felis* Bartonella henselae Bartonella quintana Bordetella parapertussis Bordetella pertussis Borrelia afzelii Borrelia burgdorferi Borrelia garinii Campylobacter coli* Campylobacter jejuni* Chlamydia pneumoniae Chlamydia psittaci Chlamydia trachomatis Haemophilus influenzae* Helicobacter pylori Klebsiella pneumoniae* Legionella bozemanii* Legionella dumoffii* Legionella gormanii* Legionella jordanis* Legionella micdadei* Legionella pneumophila serotypes 1-14 Listeria monocytogenes 1/2 a, 4b* Mycoplasma hominis Mycoplasma pneumoniae Treponema pallidum Treponema phagedenis Ureaplasma urealyticum Yersinia enterocolitica* Bacteria antigens (EUROPLUS): Borrelia VlsE (recombinant) Borrelia OspC Yeasts: Candida albicans Candida glabrata* Candida krusei* Candida parapsilosis* Candida tropicalis* Parasites: Echinococcus granulosus Leishmania donovani Plasmodium falciparum HRP-2/MSP-2 (rec.)* Plasmodium vivax MSP/CSP (recombinant)* Toxoplasma gondii Profiles: Accompanying hepatitis profile Central nervous system profile Exanthema profile Fever profile South East Asia Flavivirus profile Gastrointestinal tract profile Infectarthritis profile Infectarthritis profile (The Tropics) Lymphadenitis profile Myocarditis profile Ophthalmology profile Otitis profile Pregnancy profile Respiratory tract profile Sexually transmitted diseases (STD) profile TORCH profile * Currently not available as IVD in the EU. Special substrate combinations on request Made in Germany EUROIMMUN AG · D-23560 Luebeck (Germany) · Seekamp 31 · Phone +49 451 58550 · Fax 5855591 · E-mail [email protected] · www.euroimmun.de Test Characteristics Anti-Chlamydia trachomatis IIFT EUROIMMUN IIFT Autoimmune Diagnostics tissue/cell substrates: adrenal gland, monkey bladder, rat cartilage (trachea), monkey* cerebellum, monkey cerebrum, monkey Crithidia luciliae erythrocytes, human* eye, monkey* granulocytes, human (ethanol-fixed) granolocytes, human (formaldehyde-fixed) granulocytes, human (methanol-fixed) heart, monkey HEp-2 cells HEp-20-10 cells HUVEC hypothalamus, monkey* intestine, monkey kidney, monkey kidney, mouse kidney, rat lacrimal gland, monkey lip, monkey* liver, monkey liver, mouse liver, rat lobus temporalis, monkey* lung, monkey lymph nodes, monkey* lymphocytes, human* mamma, monkey* mouth mucosa, monkey* nerve, monkey oesophagus, monkey oesophagus, rat ovary, monkey pancreas, monkey parathyroid gland, monkey parotid gland, monkey pituitary gland, monkey placenta, monkey* prostate, monkey Saccharomyces cerevisiae skeletal muscle, monkey spermatozoa, human spleen, monkey* spinal cord, monkey stomach, monkey stomach, mouse stomach, rat synovium, monkey testis, monkey thrombocytes, human thymus, monkey thyroid gland, monkey tongue, monkey VSM47 cells umbilical cord, human EUROPLUS™ substrates: AIH (LC-1 + SLA/LP) AMA-M2 gliadin intrinsic factor myeloperoxidase (MPO) PBC (AMA M2 + Sp100) proteinase 3 (PR3) ribosomal P-proteins + Jo-1 SS-A + SS-B SS-B + ribosomal P-proteins + Jo-1 SS-B + Scl-70 + Jo-1 thyroglobulin (TG) BIOCHIP MosaicsTM: ANA global test: HEp-20-10/monkey liver Autoantibody Profile: combination of 30 different tissues per slide CIBD Profile: monkey pancreas/intest. goblet cells (culture)/ granulocytes (EOH)/Saccharomyces cerevisiae Basic Profile: HEp-20-10/monkey liver/ rat kidney/rat stomach EUROPLUS endomysium + gladin: monkey intestine/monkey liver/gliadin Granulocyte Mosaic: granulocytes (EOH)/ granulocytes (HCHO)/HEp-2/monkey liver Liver Mosaic: HEp-2/monkey liver/rat liver rat kidney/rat stomach/monkey heart Neuronal Antibody Screen: monkey cerebellum/ monkey nerve/monkey intestine Polyendocrinopathy Mosaic: monkey thyroid/ monkey pancreas/monkey adrenal/ monkey ovary/monkey testis/monkey stomach Other mosaics also available Special substrate combinations on request Test principle: The indirect immunofluorescence test is a standardized in vitro assay for the determination of specific antibodies against C. trachomatis. BIOCHIPs are coated with C. trachomatis infected cells and fixed onto the reaction fields of a microscope slide. With EUROIMMUN BIOCHIP Mosaics®, different substrates can be positioned next to each other in one reaction field and incubated with one drop of diluted serum, allowing a detailed patient antibody profile to be established with a single test. Test procedure: EUROIMMUN BIOCHIP slides are incubated using the proprietary TITERPLANE™ Technique, which enables multiple samples to be incubated next to each other and simultaneously under identical conditions. Results are evaluated by fluorescence microscopy. Incubation of the substrate with the positive and negative control sera provided in each kit verifies correct performance of the test and aids evaluation. Incubation with the TITERPLANE™ Technique Reagent tray Slide fitted with BIOCHIPs ;;; ;;;;; ;;;; ;;;; ;;;; ;; ;;;; ;;;; ;;;; ;;;; ;; ;;;; ;;;; ;;;; ;;;; ;; Inter-lot reproducibility: Inter-lot reproducibility was tested with more than 10 different lots. The deviation in the fluorescence intensity of the IIFT amounted to to no more than ± 1 intensity level for all samples. Reference range: titer 1: < 10 (IgM), titer 1: < 100 (IgA), titer 1: < 320 (IgG) The following antibody prevalences were determined for healthy blood donors (origin of samples: Germany): IgG: 22% (n = 100); IgA: 1% (n = 200); IgM: 3% (n = 200). Specificity and sensitivity: Substrate Reference (number and origin of samples) Specificity C. trachomatis (IgG) Clinical specificity (healthy blood donors, n = 100, Germany) Direct detection (n = 100, Germany) 78% C. trachomatis (IgA) Anti-Chlamydia trachomatis ELISA (n = 106, Germany) Anti-Chlamydia trachomatis ELISA (n = 32, Germany) 86% C. trachomatis (IgM) Clinical specificity (healthy blood donors, n = 200, Germany) Direct detection (n = 32, Germany) 98% Sensitivity 79% 85% 63% Technical Data: Antigen substrate Chlamydia trachomatis infected cells (species EU 70). Sample dilution Serum or plasma. Qualitative evaluation: 1:10 (IgM), 1:100 (IgA), 1:320 (IgG) Quantitative evaluation: from 1:10 (IgM), 1:100 (IgA), 1:320 (IgG) Test procedure 30 min (sample) / 30 min (conjugate), room temperature. Microscopy Objective 20x Excitation filter: 488 nm, colour separator: 510 nm, blocking filter: 520 nm Light source: EUROIMMUN LED or mercury vapour lamp, 100 W Reagents Ready for use, with the exception of the PBS-Tween buffer (for dilution and washing). Stability All kit components are stable for up to 18 months from the date of manufacture. Standard kit formats 10 or 20 slides, each containing 3, 5 or 10 test fields. Kits include all necessary reagents (except EUROSORB for the RF absorption, order no.: ZF1270-0145). Order no. FI 2191-1005 A, G or M (test kits with 10 slides, each with 5 test fields) Related products FI 2191-1005-3 G (Anti-Chlamydia MIF) * Currently not available as IVD in the EU. Made in Germany Version: 08/08 FI_2191_D_UK_B01 EUROIMMUN AG · D-23560 Luebeck (Germany) · Seekamp 31 · Phone +49 451 58550 · Fax 5855591 · E-mail [email protected] · www.euroimmun.de