Labreport The Newsletter of The Doctors Laboratory Test Update SPRING 2007 PROFILE OF 7 PCR TESTS FROM ONE SAMPLE Test Code Sexual Health Screening DL12 DL12 7 tests by PCR* from ONE of four different sample types OR REQUEST SINGLE TESTS Test Code Urine* PCR Swab Cytyc Vial Semen • First Catch Urine • Semen • Thin Prep Vial • PCR Swab N.gonorrohea Chlamydia trachomatis CT/NG combined Mycoplasma genitalium Ureaplasma urealyticum Trichomonas vaginalis Gardnerella vaginalis Herpes Simplex I/II CGON CPCR CCG MGEN UGEN TVPC GVPC HERD ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ * Polymerase chain reaction (PCR) is a molecular technique that allows a small amount of DNA to be amplified exponentially. Chlamydia trachomatis: the most common sexually transmitted infection, with Neisseria gonorrhoeae being the second most common bacterial STI in the UK. These two infections are well recognised and documented. The prevalence is highest in young sexually active adults, especially women ages 16 to 24 years and men ages 18 to 29 years. Most infections are asymptomatic, with a large proportion remaining undiagnosed. Untreated genital chlamydial infection may have long-term consequences, especially in women in whom it is a well-established cause of pelvic inflammatory disease (PID), ectopic pregnancy and infertility. Mycoplasma genitalium: has been very difficult to culture from genital tract specimens known to be positive by PCR and has been shown to be associated with nonchlamydial, nongonococcal urethritis, epididymitis and prostatitis. When men present to a clinic for treatment of persisting or recurrent urethritis after a course of doxycycline, nearly half are infected with M. genitalium. In women M. genitalium can be detected in the upper genital tract, as well being as a cause for cervicitis and urethritis – and in those with infected male partners. Ureaplasma urealyticum is similiarly identified as the most common cause of Sample Type 1 OR 1 OR 1 OR 1 x x x x Turnaround Time First Catch Urine Sample PCR Swab Thin Prep Vial Semen Sample 4 working days Turnaround Time 2 2 2 4 4 4 4 4 working working working working working working working working days days days days days days days days *First catch random urine - If requesting urine for m/c/s please ensure this is clearly requested as a separate test. symptomatic nongonococcal and nonchlamydial urethritis. (* Sex Transm Infect 2006;82:276-279) Vaginitis due to Trichomonas vaginalis is a very common sexually transmitted disease causing an offensive vaginal discharge, associated with vulvar itching, burning, redness and swelling. Testing by PCR will outperform culture. It is pathogenic to the genitourinary tract – 50% of women are asymptomatic carriers and the other 50% are symptomatic. Bacterial vaginosis (BV) formerly known as nonspecific vaginitis, is the most common cause of vaginitis and is characterised by an increase in vaginal discharge and overgrowth of certain bacteria in the vagina, including Gardnerella vaginalis. The incidence of BV increases with the number of recent and lifetime sexual partners or new sexual partners. G. vaginalis is predominantly identified in women. Male partners are usually asymptomatic and rarely develop infections with G. vaginalis – but men whose sexual partners have symptoms of BV may present with urethral colonisation of the same strain of G. vaginalis. There is sufficient data to support infection as causative agents of chronic prostatitis in men and chronic pelvic pain syndrome in women. These patients are not infrequently found to have a wide variety of bacterial infection identified by PCR testing despite previous negative cultures. Herpes Simplex Virus I/II has the characteristics of a viral, not a bacterial infection, for which there is no cure. Prevention is inherently dependent on behaviour, not treatment. Two types exist: type 1 (HSV-1) and type 2 (HSV-2). Both are closely related but differ in epidemiology. HSV-1 is transmitted chiefly by contact with infected saliva, whereas HSV-2 is transmitted sexually or from a mother's Continued on page 2 IN THIS ISSUE TDL FRONT PAGE • Sexual Health Screening STD Diagnoses at GUM clinics in the UK 1996 – 2005 NEWSLETTER INSERT Total of 1.8 million attendances at GUM clinics in the UK in 2005 – more than double the number recorded in 2002. • TDL FrostBox Service • The Body – Tumour markers at a glance TDL ANDROLOGY Year Syphilis Gonorrhoea Chlamydia Herpes All Diagnoses 1996 137 12579 35840 16811 4955151 2005 2814 19392 109958 19837 790443 TDL GENETICS 60% • Amplichip CYP450 Test % change ▲ 2,054% ▲ 54% ▲ 207% ▲ 18% ▲ Since 1996, there has been a substantial increase in diagnoses of most Sexually Transmitted Diseases (STDs) in the UK, and reports of two of the most common STDs have shown massive rises. Cases of uncomplicated gonorrhoea increased by 54% between 1996 and 2005, while genital chlamydia increased by 207%. Chlamydia has been the most commonly reported sexually transmitted disease since 2001, overtaking genital warts. This is the first time that a bacterial infection has held this title.The rapid increase in reports of bacterial STDs is probably in part due to a general deterioration in sexual health amongst young people and men who have sex with men. However, the greater acceptability of using GUM services and new campaigns to encourage testing have also made a contribution. UK Health Protection Agency, November 2006. • Male Infertility TEST UPDATES • PCA3 • Liver Fibrosis • HPV and Anal Cancer • Non Alcoholic Fatty Liver Disease (NAFLD) SERVICE UPDATES • TDL FrostBox Service TDL CONTACT • Who to ask for help Sexual Health Screening continued genital tract infection to her newborn. Clinical diagnosis may be difficult as patients do not always present with lesions. Cultures are helpful if the patients presents early enough but viral shedding only lasts a few days and often the lesion has crusted or disappeared by the time the patients is seen. Thus, a negative culture would not definitely rule out herpes. Measuring HSV DNA by PCR assay is more sensitive for detecting HSV in a lesion than a viral culture and will distinguish between HSV-1 and HSV-2. Tests can be requested as single assays, or as a group of 7 in TDL Profile DL12. ■ TDL Andrology Male Infertility I n some cases, a diagnosis of male infertility may be successfully treated allowing a couple to conceive naturally or with minimal medical assistance but some patients may have more significant disease. Treatments will continue to progress as an increasing number of couples seek fertility services. Infertility evaluation is usually considered for a couple with a history of unprotected intercourse for at least 12 months without a pregnancy. This is relevant even for couples who have had a successful first pregnancy but are unsuccessful establishing a second pregnancy. A full screening evaluation for the male partner incorporates a complete medical, sexual, family and social history, together with a physical examination, laboratory evaluation and possibly up to two or more semen analyses. Semen profiles can show considerable variation from one sample to the next. If one test gives poor results, it is essential to carry out a second test between 1 and 3 weeks later. It is well recognised that patients presenting with a subfertile semen analysis may later show parameters within the normal reference range on a subsequent test, thus averting an incorrect diagnosis of infertility. Alternatively, men who have had a very satisfactory semen analysis more than 6 months previously, but who are finding it difficult to conceive, may benefit from a repeat test as conditions such as primary testicular failure or seminoma may manifest with a dramatic loss in sperm concentration. ■ • • • • Test Update PCA3 The first molecular diagnostic assay that can contribute to the diagnosis of prostate cancer. D iagnosis of prostate cancer has relied heavily on serum Prostate Specific Antigen (PSA), physical examination by digital rectal examination (DRE) and biopsy. However all have shortcomings. Nonmalignant conditions (eg prostatitis and benign prostatic hyperplasia) can cause PSA levels to rise and the positive predictive value (PPV) for diagnosing cancer by serum PSA is low. Approximately 75% of men suspected of having cancer based on PSA testing actually have non‐cancerous conditions. The PPV by DRE is even lower (10%). Complications caused by prostate biopsy are well documented. As many as 10 – 25% of patients with a negative biopsy may have prostate cancer. PCA3 is prostate cancer specific, over expressed in >95% of malignant tissue, and is significantly up-regulated (60-100 fold) in prostate cancer. PCA3 is more prostate cancer specific than PSA with a positive predictive value to diagnose prostate cancer being almost twice than of serum PSA. PCA3 is only expressed in prostate tumour – not in benign or normal tissue and is not affected by the size of the prostate. Testing for PCA3 is from a urine sample. The urine sample must be collected following a DRE. The released prostate cells are collected in a first catch urine after the DRE. Gen-Probe urine sample collection tubes must be used and there are important storage and temperature instructions once the urine sample has been collected. The PCA3 result is reported as a score; the higher the score the greater the probability of a positive prostate biopsy whilst the lower the score the likelihood of a positive biopsy decreases. The greatest diagnostic utility occurs at a cut off of 35. Highly specific to Prostate Cancer Non-invasive Improves diagnosis of Prostate Cancer Helps reduce the number of unnecessary biopsies By measuring the expression of mRNA from the PCA3 gene, this new assay provides greater accuracy: a positive result may indicate prostate cancer and the need for an additional biopsy; a negative result even though accompanied by a PSA greater than 4 ng/ml suggests a relatively longer time interval may be acceptable between biopsies. The routine use of PCA3 in men with raised PSA levels with negative biopsies would expect to both enhance the early detection of cancer in these men, as well as reduce the number of unnecessary biopsies. The use of PCA3 in combination with current PSA testing and increased accuracy for timing of biopsies would also increase the biopsy yield of positive cancers (UROLOGY.2007;69:532 – 535). For further information about PCA3, or to order Gen-Probe PCA3 Urine Specimen Transport Tubes please contact Annette Wilkinson or Lynn DerbyLewis at The Doctors Laboratory on 020 7307 7373 or by email [email protected] ■ PCA3 Score As the PCA3 score increases the likelihood for positive biopsy increases. As the PCA3 score decreases, the likelihood for a positive biopsy decreases. The greatest diagnostic utility occurs at a cut-off of 35. 125 115 105 95 85 75 65 55 45 Increased Probability of Positive Biopsy 35 30 25 20 15 10 9 8 7 6 5 4 Decreased Probability of Positive Biopsy Sample information – See TDL Laboratory Guide page 56 (Tumour Markers) Test Code Sample Type Turnaround Time PCA3 PCA3 1 x Gen Probe PCA3 Sample Transport Tube 5-7 working days DRE and Urine sample collection is by appointment (020 7307 7383) at Patient Reception, 55 Wimpole Street, London W1G 8YL Sample information New Andrology Suite Test Code Instructions for Appointments Comprehensive Semen Analysis for Fertility SPER Post Vasectomy PVAS Semen Analysis Sperm DNA SEXT Please make, or ask your patient to make, an appointment for a semen analysis (Fertility or Post Vasectomy) on 0207 025 7940 or 020 7307 7373. This will ensure that samples are assessed within specific time limits. British Andrology Society guidelines currently specify that first appointments for post vasectomy samples should be at least 16 weeks and 24 ejaculations after surgery. Important information It is essential that patients do not abstain for longer than 5 days but not less than 2 days. If you would like to discuss a patient’s results, or specialist services, (Sperm Preparation for overnight survival or IUI sperm motility and vitality testing, retrograde ejaculation etc.) or would like further information about TDL Andrology’s services, please call Dr Sheryl Homa, Head of Andrology, on 020 7025 7940 or 07813 077797 or email [email protected]. TDL’s new Andrology Laboratory, with three dedicated sample collection rooms, is now located in the same premises as Patient Reception at 55 Wimpole Street, providing diagnostic semen analysis and post vasectomy analysis testing services. The laboratory is CPA accredited, and performs to current WHO, Associated Biomedical Andrologists and British Andrology Society guidelines for laboratories undertaking comprehensive semen analyses for fertility, post-vasectomy semen analyses and screening of sperm donors. TDL Andrology contributes to the external quality assurance scheme UKNEQAS. TDL Genetics Amplichip CYP450 Test Breast Cancer and Tamoxifen response T he Amplichip CYP450 Test (Roche Diagnostics) has been designed to determine a patient’s CYP2D6 and CYP2C19 genotype and phenotype. Genes such as these have a marked impact on response rates to certain drugs, helping to explain why some patients respond well, others poorly, and why some patients need higher or lower doses. The Amplichip test helps clinicians determine whether patients process drugs too slowly (poor metabolisers) or too quickly (extensive or ultrarapid metabolisers) and enables the most suitable drugs and dosages to be determined before treatment begins. Recent studies have shown that Tamoxifen may not work as well in women with breast cancer who carry certain variants of the CYP2D6 gene. An Test Update HPV and Anal Cancer H igh-risk HPV types are implicated in a substantial portion of anal, penile, and head and neck precancers and cancers. HPV 16 and 18 are the most common HPV types found in cervical cancer and are responsible for approximately 70% of these cancers. Anal cancer rates in the UK have risen by 2.5 – 5 fold in the last 50 years. They are similar to the incidence rate of other external anogenital malignancy but certain subsets of the population have a high susceptibility to anal cancer:1 • Men and women with HIV infection • Men who have sex with men Anal cancer shows many similarities to cervical cancer: high risk HPV infection being the causative factor in most cases. Rates of High Risk HPV in anal cancer specimens are 83% men / 95% women and with further advances in detection and Test Update Liver Fibrosis L iver Fibrosis with progression to cirrhosis is a common outcome in chronic liver disease, Diagnosis of liver fibrosis and monitoring to identify the effectiveness of drug treatment is usually made by histological analysis, the “reference standard” index of liver fibrosis. Histology can be highly informative but there are well documented disadvantages for undertaking biopsies – they are relatively expensive, and may not always give accurate results because of difficulties in sampling or interpretation. Fibrosis is also not evenly distributed throughout the liver and small amounts of biopsy material may give a misclassified result. Repeated liver biopsies are not considered acceptable, even through the indications for obtaining a liver biopsy for diagnosis, determining prognosis, and evaluating the impact of treatment are estimated 7% to 10% of Caucasian women with breast cancer carry such CYP2D6 variants, reducing their ability to metabolise Tamoxifen, increasing a risk of recurrence and reducing disease-free survival rates.1 The CYP2D6 enzyme transforms Tamoxifen into the potent anti-estrogen endoxifen, but certain CYP2D6 genetic variants and inhibitors of the enzyme markedly reduce endoxifen plasma concentrations in Tamoxifen-treated patients.1,2 CYP2D6 metabolism, as measured by genetic variation and enzyme inhibition, has therefore been proposed as an independent predictor of breast cancer outcome in post-menopausal women receiving tamoxifen for early breast cancer. Determination of CYP2D6 genotype may also be of value in selecting adjuvant hormonal therapy and it appears CYP2D6 inhibitors should be avoided in Tamoxifen-treated women. The effects of reduced metabolism appeared greatest in poor metabolisers, with a greater than threefold higher risk of recurrence compared to extensive metabolisers. In light of this recent evidence the FDA was advised, by its own panel of experts, to consider changing the Tamoxifen label to include a reference to CYP2D6 and the risk associated with certain metaboliser phenotypes. The results of a patient’s genotype and predicted phenotype (poor, intermediate, extensive, ultrarapid) are presented in her report and may improve the choice of optimal hormonal therapy for the treatment of estrogen receptor positive cancer. ■ 1. Goetz et al J Clin Oncol. 2005;23(36):9312-8. 2. Jin Y et al: J Natl Cancer Inst 97:30, 2005 Sample information Test Code Sample Type Turnaround Time Roche Amplichip CHIP EDTA whole blood 1-2 weeks For further information please contact Dr Lisa Levett, TDL Genetics: [email protected] or 020 7307 7409. classification, this may increase. Type16 is the most frequent HPV type detected followed by Type18, regardless of gender. The types being identified (16, 18, 31, 33, 45) are identical to the subtypes found in cervical cancer2 Smokers (men and women) are at particularly high risk for anal cancer, independent of age and other risk factors3. Anal cytology is the candidate test for screening for prevention of anal cancer. Inspection and clinical examination includes cytological, perianal and intraanal PAP smear sampling. Simultaneous HPV DNA by PCR testing increases the sensitivity of anal cytology andby using Liquid Based Cytology both cytological examination and HPV testing can be undertaken. Test values and screening performances are well documented5,6. Triage of atypical cytology can be performed according to gynaecological cancer screening procedures. If HPV infection is identified, identifying low from high risk subtypes is helpful. Biopsy, excision and examination especially for HPV lesions is recommended. Follow up testing of HPV DNA by PCR will identify whether there is clearance or persistence of the virus. Persistence of HPV infection is, like cervical cancer, indicative of an increased risk of recurrence4. ■ 1 2 3 4 5 6 Science to Community Clinical #3 J Clin Pathol 55:244 – 255 Cancer. 2004 Jul 15;101(2):270-80 AIDS 1998;12:495-503 Cancer Epidemiol Biomarkers Prev 2003;12:638-42 Colorectal Dis (2006) 21: 135 – 142 Sample information Test Code Sample Type Turnaround Time Anal Pap Smear HPV 20 subtypes* DNA by PCR APAP HP20 1 x Cytyc Thin Prep Vial 1 x PCR Swab or 1 x Thin Prep Vial 48 hours 4 days *Each of the 20 DNA subtypes are reported individually as Positive or Negative (5 Low and 15 High Risk). This test is appropriate for male or female patients. important. Whether the damage is caused by alcohol, fat or virus the effect is much the same - long term inflammation causing scar formation or fibrosis – with scar tissue eventually obstructing blood flow. This is cirrhosis. Many chronic liver diseases can now be diagnosed without resorting to biopsy, with the assessment of liver inflammation made by using blood tests. Because Enhanced Liver Test Function (ELF TM) test uses a blood sample rather than a biopsy, and it can be used repeatedly for the same patient helping monitor progression or reduction of fibrosis. The Enhanced Liver Fibrosis (ELF TM) test combines three direct serum markers and employs a proprietary algorithm* based on the measurement of three key markers: hyaluronic acid, TIMP-1 and PIIINP. The algorithm measures each of these markers by immunoassay, to create an ELF TM score able identify patients with little or no fibrosis, from clinically significant hepatic fibrosis. ELF TM testing can be used to monitor disease progression and response to therapy and can be used in many chronic conditions, including alcoholic liver disease, primary biliary cirrhosis, and hepatitis. ■ * Non-invasive markers of liver fibrosis tested in different forms of chronic liver disease showed variable diagnostic performance, but accuracy was rarely above 75%-80%. Results obtained when markers are combined (the set of algorithms that combine sequentially indirect non-invasive markers of liver fibrosis) achieve 90%-95% diagnostic accuracy. The accuracy of this algorithm is similar when compared to three expert liver pathologists working independently. Sample information Test Code Sample Type Turnaround Time Enhanced Liver Fibrosis (ELF) Test ELF 2 x Gold/SST 7 days This test employs an algorithm based on the measurement of three key direct liver markers: hyaluronic acid, TIMP-I and PIIINP*. This serum sample needs no special handling. Postal packs are routinely provided on request by The Doctors Laboratory. Test Update Non Alcoholic Fatty Liver Disease (NAFLD) W ith an estimated prevalence as high as 30% in Western countries, NAFLD is emerging as one of the commonest causes of abnormal liver function. The effect of obesity in affluent countries means that it has overtaken alcohol and viral infection as the main cause of liver disease. The effects of excess fat has only recently started to be recognised. Although its true prevalence is unknown, some estimates suggest it may already affect up to one-third of adults. Diagnosis is not easy. Most subjects with NAFLD are asymptomatic but it has a well-established capability of leading to cirrhosis and end-stage liver disease in affected patients. It is not clear why an excess of fat harms the liver – most people with liver disease do not look like they have liver disease. Because liver biopsy remains the gold standard for grading and staging (ie, recognition of inflammation and fibrosis), a critical issue in management is whether individual patients should undergo biopsy to detect progressive disease. Biopsies have inherent drawbacks – they are invasive, relatively expensive, and small amounts of biopsy material may not be representative – and for this reason referral for liver biopsy is likely to be considered only after a thorough, non-invasive serologic and radiographic evaluation have failed to confirm a diagnosis of cirrhosis; the benefit of biopsy outweighs the risk; and it is postulated that biopsy will have a favorable impact on the treatment of chronic liver disease. Based on available evidence, it can be anticipated that non-invasive markers of liver fibrosis and their combined use will become a most useful tool in the clinical management of many forms of chronic liver disease. However, their implementation will not completely eliminate the need for liver biopsy. The Enhanced Liver Fibrosis (ELFTM) test is validated as highly accurate across all stages of disease severity in HCV, HIV, HCV/HIV co-infection, primary biliary cirrhosis and non alcoholic fatty liver disease. ■ Sample information Test Code Sample Type Turnaround Time Enhanced Liver Fibrosis (ELF) Test ELF 2 x Gold/SST 7 days This test employs an algorithm based on the measurement of three key direct liver markers: hyaluronic acid, TIMP-I and PIIINP*. This serum sample needs no special handling. Postal packs are routinely provided on request by The Doctors Laboratory. Service Update Sample information TDL FrostBox Service S ome tests need special handling or need to be frozen at the time of sample taking. Without regular deliveries of dry ice, making arrangements for same day deliveries are expensive and time consuming. The TDL FrostBox Service is a one price solution for anyone within the UK who wants to arrange regular, or ad hoc, shipments for one or more frozen samples. We provide a simple, cost effective, door to door total service at the times you need it. Test Code Instructions Next day delivery & collection TDL Frostbox FBOX Telephone 020 7307 7380 or email [email protected] £97.00 If you have an on-site freezer, take your sample, prepare and freeze at your convenience – store as frozen until your FrostBox arrives. If you do not have a freezer wait until your Frost Box arrives before taking and preparing the sample. Label samples with their three unique identifiers, include the request form(s) or work sheets in the sample bag(s) and surround with dry ice. Handle dry ice in accordance with health and safety guidelines. Dry ice should not be put into food or drinks. ■ 1 4 Deliver at TDL on Wednesday 3 24 hour Telephone (Main Switchboard for all services) 020 7307 7373 Fax 020 7307 7374 David Byrne [email protected] Group Laboratory Director Rob Joyce [email protected] Director of Sales/Service Annette Wilkinson [email protected] Director of TDL Genetics Dr Lisa Levett [email protected] Heads of Support Departments Quality Management Cyril Taylor [email protected] Customer Service Manager Rochelle Fakhri [email protected] Patient/Doctor Invoices Spencer Jack [email protected] Courier Control Steven Sykes [email protected] Patient Reception/Home Visits Michaela Rackham [email protected] Pathology Supplies Ken Roberts [email protected] Service Support Sample Handling Jacqui Dehaney [email protected] Call Centre Adrian Worley [email protected] IT Alan Stevens [email protected] TDL FrostBox 2 Arrive Tuesday Pick up Tuesday TDL Contact: Who to ask for help CEO Order from TDL on Monday The TDL Frostbox process: • Your FrostBox will arrive by 10.00am • Your FrostBox will be collected by 4.00pm on the same day. • Your FrostBox will be delivered to Sample Reception at TDL by 9.00am the following day. Order your TDL FrostBox by telephone: 020 7307 7380 or by email: [email protected], giving details of: • Who you are • Where the TDL FrostBox needs to be delivered Heads of Laboratory Departments Sample Handling and Data Entry Kathleen Frost [email protected] Referrals Lynn Derby-Lewis [email protected] Haematology Letisha Brukman [email protected] Biochemistry Barbara Vale [email protected] Microbiology Massimo Bonaiti [email protected] Andrology Dr Sheryl Homa [email protected] Cytology Nick McClenaghan [email protected] Immunology/Virology Nalin Shah [email protected] Molecular Genetics Dr Stuart Liddle [email protected] Cytogenetics Terry Ballard [email protected] TDL Trials Abraham Roodt [email protected] TAP506/24-04-07/V7 • Who is your contact person • Your direct telephone number • Which day/date you want your FrostBox to be delivered (TDL FrostBoxes can be delivered and collected on Mondays within the M25 only). Orders for a Friday FrostBox, for return delivery to the laboratory on Saturday morning will incur an additional charge of £20.00) • Please order by 2.00pm for a next day delivery The charge for this service is £97.00 and will be invoiced with your pathology.
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