First-catch Urine Specimens Research Articles

Self-collected glans/meatal 'dry' swab specimen and NAAT technology detects Chlamydia trachomatis and Neisseria gonorrhoeae - implications for public policy changes.

Increasing Chlamydia trachomatis (CT) rates and ever-present Neisseria gonorrhoeae (NG) infections in women have given rise in the past to consideration of male screening programs in order to address the silent male reservoir. Non-medical venues (e.g. home collection, restrooms or other private locations) may be viable venues to reach certain populations that in the past have not been accessed. Effortlessly collected, non-invasive, self-collected male specimens that are stable and easy to transport would enhance the success of male screening programs. We designed a head-to-head study to consider the effectiveness of non-invasive self-collected glans/meatal dry swab (SCS) specimens to detect CT and NG nucleic acid when compared to traditional clinician-collected swab (CCS) specimens and first-catch urine (FCU) specimens. A total of 284 male patients were included in the study. Specimens were processed using the Becton Dickinson ProbeTec ET system. The overall sensitivity of SCS was 91.1% with a specificity of 99.2%. There was an overall SCS agreement of 97.7% with CCS specimens and 90.4% with FCU specimens. Dry swab specimens are easy to collect, transport and test. Non-invasive dry self-collected glans/meatal swab specimens are a viable specimen choice.

A non-invasive urine exosome gene expression assay (ExoIntelliScore Prostate) to predict pathologic stage and grade in the prostatectomy specimen.

46 Background: Non-invasive screening tools that add predictive value for identifying high-grade, Gleason score (GS) ≥7 should impact on the current diagnostic and subsequent treatment decision process. We have developed standardized processes to isolate exosomal-RNA from first-catch, non-DRE, urine specimens and have validated a three gene signature (ExoIntelliScore Prostate) that reliably differentiates GS7+ from GS6 and benign at the time of an initial biopsy. In addition to the biopsy outcomes, we sought to understand the performance of the assay in predicting objective pathology in the prostatectomy specimen. Methods: Pre-radical prostatectomy first-catch non-DRE urine specimens were collected from 5 urology practices (academic/community practices) without preservative, stored at 2-8C (up to 2 weeks) and shipped on ice to a central laboratory (Exosome Diagnostics, St. Paul, MN). Exosomes were isolated and RNA extraction performed. RT-qPCR RNA copy numbers of ERG and PCA3 were normalized to SPDEF. Spearman and Pearson correlation along with AUC was used to evaluate performance. Results: Pre-RP urine samples from 430 patients were collected: 16% GS6, 45% GS7 (3+4), 39% (>/=GS4+3) and 85% PSA <10 ng/mL. 359 patients had complete clinical and ExoIntelliScore Prostate assay results with comparable demographics. Importantly, the Exo106 significantly correlated with prostatectomy GS (p=0.025; Spearman’s rank-order). In addition, Exo106 significantly correlated with both tumor volume and pathologic stage (p=0.002, Pearson). Conclusions: A previously validated non-invasive urine exosome gene signature (ExoIntelliScore Prostate) accurately reflects what is present within the prostatectomy specimen with respect to grade, stage and tumor volume. Such information could be incorporated into the treatment decision process, including active surveillance.

A first catch, non-DRE urine exosome gene signature to predict Gleason 7 prostate cancer on an initial prostate needle biopsy.

45 Background: There is a critical need to reduce initial prostate biopsies (PB) in the PSA ‘gray zone’ of 2-10 ng/mL. Non-invasive screening tools that add predictive value for identifying high-grade, Gleason score (GS) ≥7 should impact on the current diagnostic paradigm. We have developed standardized processes to isolate exosomal-RNA from first-catch urine specimens and sought to identify a gene signature that reliably differentiates GS7+ from GS6 and benign disease. Methods: First-catch non-DRE urine specimens were collected at six sites in standard collection vessels without preservative, stored at 2-8C (up to two weeks) and shipped on ice to a central laboratory (Exosome Diagnostics, St. Paul, MN). Upon receipt, samples were filtered (0.8um), and exosome isolation and RNA extraction performed. RT-qPCR RNA copy numbers of ERG and PCA3, normalized to SPDEF, were measured to generate a three-gene signature, defined to yield a score S between 0 and 30, where S>10 predicts GS7+ vs. GS6 and benign lesions with optimal NPV, Sensitivity and Specificity. Results: Urine samples from 170 men (PSA 2-10 ng/mL, first biopsy,40 mL; median age 62 years, median PSA 5.1 ng/mL, 70% negative DRE, 77%, no family history, 82% Caucasian) of 453 total, with comparable demographics, were evaluated. With 46% and 21% prevalence of positive biopsy for any cancer or GS7+ disease, respectively, a dichotomous gene signature demonstrated good clinical performance in predicting biopsy result. For GS7+, the NPV and PPV were 98.6% and 34.7%, respectively. A continuous score alone had an AUC of 0.76 for discriminating GS7+ from GS6 and benign disease and the results were significantly better than the prostate cancer prevention trial risk calculator (PCPTRC), AUC 0.60. Conclusions: A novel, non-invasive urine exosome gene signature demonstrated independent, negative predictive value for the diagnosis of GS7+ from first biopsy patients with ‘gray zone’ serum PSA. Its use in the biopsy decision process could result in fewer prostate biopsies once validated on an expanded prospective cohort.

A novel urinary long non-coding RNA transcript improves diagnostic accuracy in patients undergoing prostate biopsy.

Long non-coding RNA (LncRNA) PCA3 has been a well-established urine biomarker for the detection of prostate cancer (PCa). Our previous study showed a novel LncRNA FR0348383 is up-regulated in over 70% of PCa compared with matched benign tissues. The aim of this study was to evaluate the diagnostic value of urinary FR0348383 for men undergoing prostate biopsy due to elevated PSA (PSA > 4.0 ng/ml) and/or abnormal digital rectal examination (DRE). Post-DRE first-catch urine specimens prior to prostate biopsies were prospectively collected. After the whole transcriptome amplification, quantitative real time polymerase chain reaction was applied to quantify urine FR0348383 and PSA levels. The FR0348383 score was calculated as the ratio of PSA and FR0348383 mRNA (PSA mRNA/FR0348383 mRNA × 1000). The diagnostic value of FR0348383 score was evaluated by logistic regression and decision curve analysis. 213 cases with urine samples containing sufficient mRNA were included, 94 cases had serum PSA level 4.0-10.0 ng/ml. PCa was identified in 72 cases. An increasing FR0348383 score was correlated with an increasing probability of a positive biopsy (P < 0.001). Multivariable logistic analysis indicated FR0348383 score (P < 0.001), PSA (P = 0.004), age (P = 0.007), prostate volume (P < 0.001) were independent predictors of PCa. ROC analysis demonstrated FR0348383 score outperformed PSA, %free PSA, and PSA Density in the prediction of PCa in the subgroup of patients with grey area PSA (AUC: 0.815 vs. 0.562 vs. 0.599 vs. 0.645). When using a probability threshold of 30% in the grey zone cohort, The FR0348383 score would save 52.0% of avoidable biopsies without missing any high grade cancers. FR0348383 transcript in post-DRE urine may be a novel biomarker for detection of PCa with great diagnostic value, especially in the grey zone cohort. The application of FR0348383 score in clinical practice might avoid unnecessary prostate biopsies and increase the specificity of PCa diagnosis.

Use of the UriSwab collection device for testing of Chlamydia trachomatis and Neisseria gonorrhoeae: implications for a postal testing service

In order to demonstrate the reliability of UriSwab, a trial was conducted using urine samples that had previously returned a detected result for Chlamydia trachomatis and/or Neisseria gonorrhoeae. Urine specimens (115 samples) were received from sexual health clinics and tested using the Roche Cobas 4800 CT/NG method. Concurrently, the urine samples were pipetted directly on to the sponge applicator of the UriSwab, simulating micturition, and the urine harvested from the UriSwab was tested using the Roche Cobas 4800 method. Of the 87 standard urine specimens that were C. trachomatis detected, 85 (98%) were also detected in the corresponding UriSwab specimen (sensitivity 97.7%, specificity 95.7%). Of the 34 standard specimens that were N. gonorrhoeae detected, 33 (97%) were also detected in the corresponding UriSwab specimen (sensitivity 97.1%, specificity 100%). The performance of the UriSwab in this trial was comparable with the testing of neat first-catch urine specimens for both C. trachomatis and N. gonorrhoeae.

Prospective Multicentre Evaluation of PCA3 and TMPRSS2-ERG Gene Fusions as Diagnostic and Prognostic Urinary Biomarkers for Prostate Cancer

BackgroundProstate cancer antigen 3 (PCA3) and v-ets erythroblastosis virus E26 oncogene homolog (TMPRSS2-ERG) gene fusions are promising prostate cancer (PCa) specific biomarkers that can be measured in urine. ObjectiveTo evaluate the diagnostic and prognostic value of Progensa PCA3 and TMPRSS2-ERG gene fusions (as individual biomarkers and as a panel) for PCa in a prospective multicentre setting. Design, setting, and participantsAt six centres, post–digital rectal examination first-catch urine specimens prior to prostate biopsies were prospectively collected from 497 men. We assessed the predictive value of Progensa PCA3 and TMPRSS2-ERG (quantitative nucleic acid amplification assay to detect TMPRSS2-ERG messenger RNA [mRNA]) for PCa, Gleason score, clinical tumour stage, and PCa significance (individually and as a marker panel). This was compared with serum prostate-specific antigen and the European Randomised Study of Screening for Prostate Cancer (ERSPC) risk calculator. In a subgroup (n=61) we evaluated biomarker association with prostatectomy outcome. Outcome measurements and statistical analysisUnivariate and multivariate logistic regression analysis and receiver operating curves were used. Results and limitationsUrine samples of 443 men contained sufficient mRNA for marker analysis. PCa was diagnosed in 196 of 443 men. Both PCA3 and TMPRSS2-ERG had significant additional predictive value to the ERSPC risk calculator parameters in multivariate analysis (p<0.001 and resp. p=0.002). The area under the curve (AUC) increased from 0.799 (ERSPC risk calculator), to 0.833 (ERSPC risk calculator plus PCA3), to 0.842 (ERSPC risk calculator plus PCA3 plus TMPRSS2-ERG) to predict PCa. Sensitivity of PCA3 increased from 68% to 76% when combined with TMPRSS2-ERG. TMPRSS2-ERG added significant predictive value to the ERSPC risk calculator to predict biopsy Gleason score (p<0.001) and clinical tumour stage (p=0.023), whereas PCA3 did not. ConclusionsTMPRSS2-ERG had independent additional predictive value to PCA3 and the ERSPC risk calculator parameters for predicting PCa. TMPRSS2-ERG had prognostic value, whereas PCA3 did not. Implementing the novel urinary biomarker panel PCA3 and TMPRSS2-ERG into clinical practice would lead to a considerable reduction of the number of prostate biopsies.

Bacterial communities of the coronal sulcus and distal urethra of adolescent males.

Lactobacillus-dominated vaginal microbiotas are associated with reproductive health and STI resistance in women, whereas altered microbiotas are associated with bacterial vaginosis (BV), STI risk and poor reproductive outcomes. Putative vaginal taxa have been observed in male first-catch urine, urethral swab and coronal sulcus (CS) specimens but the significance of these observations is unclear. We used 16 S rRNA sequencing to characterize the microbiota of the CS and urine collected from 18 adolescent men over three consecutive months. CS microbiotas of most participants were more stable than their urine microbiotas and the composition of CS microbiotas were strongly influenced by circumcision. BV-associated taxa, including Atopobium, Megasphaera, Mobiluncus, Prevotella and Gemella, were detected in CS specimens from sexually experienced and inexperienced participants. In contrast, urine primarily contained taxa that were not abundant in CS specimens. Lactobacilllus and Streptococcus were major urine taxa but their abundance was inversely correlated. In contrast, Sneathia, Mycoplasma and Ureaplasma were only found in urine from sexually active participants. Thus, the CS and urine support stable and distinct bacterial communities. Finally, our results suggest that the penis and the urethra can be colonized by a variety of BV-associated taxa and that some of these colonizations result from partnered sexual activity.

2105 ERG IMMUNOEXPRESSION IS NOT ASSOCIATED WITH INCREASED RISK OF RECURRENCE AFTER PROSTATECTOMY FOR CLINICALLY-LOCALIZED PROSTATE CANCER: A NESTED CASE CONTROL STUDY

post-DRE first-catch urine specimens were collected prior to prostate biopsy. Progensa PCA3 and T2:ERG (quantitative nucleic acid amplification assay to detect T2:ERG mRNA) were assessed and correlated (individually and as a marker panel) to prostate cancer diagnosis, Gleason score, and clinical T stage. This was compared to serum PSA. In a subgroup (n 61) prostatectomy outcome was evaluated. The independent sample T-test and the non-parametric Mann Whitney test were used. RESULTS: Of the 497 men that were included, urine samples of 443 men contained sufficient mRNA for marker analysis. Table 1 shows the diagnostic and prognostic values of the markers. Prostate cancer was diagnosed in 196/443 men. Serum PSA, PCA3 and T2: ERG were all significantly correlated with diagnosis prostate cancer. PCA3 using cutoff 25 had the strongest correlation (p 6.3 10-13), exceeding serum PSA. By combining PCA3 and T2:ERG the correlation improved significantly (p 5.4 10-17). The AUC (area under the curve) increased from 0.720 (only PCA3) to 0.760 for the marker panel PCA3 T2:ERG. The sensitivity increased from 68% to 76%. T2:ERG was significantly correlated with biopsy Gleason score, clinical T stage, and extracapsular extension. PCA3 had no correlation with these prognostic parameters. CONCLUSIONS: PCA3 is a strong diagnostic marker for prostate cancer. By combining PCA3 and T2:ERG the diagnostic value and the AUC increases evidently. In this prospective multicenter study T2:ERG had significant prognostic value, there was no correlation of PCA3 with Gleason score and T stage.

2104 PROSPECTIVE MULTICENTER EVALUATION OF PCA3 AND TMPRSS2-ERG GENE FUSIONS AS DIAGNOSTIC AND PROGNOSTIC BIOMARKERS FOR PROSTATE CANCER

You have accessJournal of UrologyProstate Cancer: Markers I1 Apr 20122104 PROSPECTIVE MULTICENTER EVALUATION OF PCA3 AND TMPRSS2-ERG GENE FUSIONS AS DIAGNOSTIC AND PROGNOSTIC BIOMARKERS FOR PROSTATE CANCER Gisele Leyten, Daphne Hessels, Erik Cornel, Theo de Reyke, Henk Vergunst, Paul Kil, Ben Knipscheer, Sander Jannink, Frank Smit, Hans de Jong, Inge van Oort, Peter Mulders, and Jack Schalken Gisele LeytenGisele Leyten Nijmegen, Netherlands More articles by this author , Daphne HesselsDaphne Hessels Nijmegen, Netherlands More articles by this author , Erik CornelErik Cornel Hengelo, Netherlands More articles by this author , Theo de ReykeTheo de Reyke Amsterdam, Netherlands More articles by this author , Henk VergunstHenk Vergunst Nijmegen, Netherlands More articles by this author , Paul KilPaul Kil Tilburg, Netherlands More articles by this author , Ben KnipscheerBen Knipscheer Emmen, Netherlands More articles by this author , Sander JanninkSander Jannink Nijmegen, Netherlands More articles by this author , Frank SmitFrank Smit Nijmegen, Netherlands More articles by this author , Hans de JongHans de Jong Nijmegen, Netherlands More articles by this author , Inge van OortInge van Oort Nijmegen, Netherlands More articles by this author , Peter MuldersPeter Mulders Nijmegen, Netherlands More articles by this author , and Jack SchalkenJack Schalken Nijmegen, Netherlands More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2012.02.2272AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES In 2007 it was published that the combined use of urine markers PCA3 and TMPRSS2-ERG gene fusions (T2:ERG) increased diagnostic sensitivity for prostate cancer. The aim of this study was to evaluate the diagnostic and prognostic value of Progensa PCA3 and T2:ERG (as individual biomarkers and as a panel) in a prospective multicenter setting. METHODS Prospective multicenter study (6 clinics) in which post-DRE first-catch urine specimens were collected prior to prostate biopsy. Progensa PCA3 and T2:ERG (quantitative nucleic acid amplification assay to detect T2:ERG mRNA) were assessed and correlated (individually and as a marker panel) to prostate cancer diagnosis, Gleason score, and clinical T stage. This was compared to serum PSA. In a subgroup (n=61) prostatectomy outcome was evaluated. The independent sample T-test and the non-parametric Mann Whitney test were used. RESULTS Of the 497 men that were included, urine samples of 443 men contained sufficient mRNA for marker analysis. Table 1 shows the diagnostic and prognostic values of the markers. Prostate cancer was diagnosed in 196/443 men. Serum PSA, PCA3 and T2:ERG were all significantly correlated with diagnosis prostate cancer. PCA3 using cutoff 25 had the strongest correlation (p=6.3×10-13), exceeding serum PSA. By combining PCA3 and T2:ERG the correlation improved significantly (p=5.4×10-17). The AUC (area under the curve) increased from 0.720 (only PCA3) to 0.760 for the marker panel PCA3+T2:ERG. The sensitivity increased from 68% to 76%. T2:ERG was significantly correlated with biopsy Gleason score, clinical T stage, and extracapsular extension. PCA3 had no correlation with these prognostic parameters. CONCLUSIONS PCA3 is a strong diagnostic marker for prostate cancer. By combining PCA3 and T2:ERG the diagnostic value and the AUC increases evidently. In this prospective multicenter study T2:ERG had significant prognostic value, there was no correlation of PCA3 with Gleason score and T stage. © 2012 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 187Issue 4SApril 2012Page: e848-e849 Advertisement Copyright & Permissions© 2012 by American Urological Association Education and Research, Inc.MetricsAuthor Information Gisele Leyten Nijmegen, Netherlands More articles by this author Daphne Hessels Nijmegen, Netherlands More articles by this author Erik Cornel Hengelo, Netherlands More articles by this author Theo de Reyke Amsterdam, Netherlands More articles by this author Henk Vergunst Nijmegen, Netherlands More articles by this author Paul Kil Tilburg, Netherlands More articles by this author Ben Knipscheer Emmen, Netherlands More articles by this author Sander Jannink Nijmegen, Netherlands More articles by this author Frank Smit Nijmegen, Netherlands More articles by this author Hans de Jong Nijmegen, Netherlands More articles by this author Inge van Oort Nijmegen, Netherlands More articles by this author Peter Mulders Nijmegen, Netherlands More articles by this author Jack Schalken Nijmegen, Netherlands More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Sexually Transmitted Diseases: Office-Based Screening and Diagnostic Tests

Screening and diagnostic testing are crucial for preventing the sequelae and ongoing transmission of sexually transmitted diseases (STDs). Several tests are available; relevant terms and abbreviations are presented in the table. Chlamydia trachomatis and Neisseria gonorrhoeae Annual chlamydial screening in sexually active women younger than 26 is vital for preventing pelvic inflammatory disease, ectopic pregnancy, and infertility associated with this often asymptomatic infection.1 Nucleic acid amplification testing (NAAT) is the most sensitive method for diagnosing chlamydia and gonorrhea. Vaginal swabs identify more chlamydial infections than cervical swabs or first-catch urine specimens, perhaps because swabbing the vagina collects material from both the cervix and urethra.2 Vaginal swab collection does not require a speculum, and patient-collected vaginal swabs are as sensitive as clinician-collected specimens.2 Many women prefer self-collection of vaginal swabs over urine collection.3 NAAT can also be performed on liquid-based cytology specimens, but the sensitivity of this approach may be lower than that of NAAT in cervical swabs.4 Antibiotic-Resistant Neisseria gonorrhoeae The proportion of gonococcal isolates …

P3-S1.30 Successful use of non-invasive self obtained glans/meatal dry FLOQSwabs (COPAN) for CT/NG detection with the BD ProbeTec ET assay in Northern Italy

BackgroundIncreasing Chlamydia trachomatis (CT) rates and ever-present Neisseria gonorrhoeae (NG) infections worldwide in women have led to the consideration of targeted male CT/NG screening programs in order to address the...

P3-S1.31 Comparison of Copan UriSwab with BD ProbeTec urine preservative transport kit for preservation and detection of CT and NG in the ProbeTec assay

BackgroundUrine specimen collection is better accepted by the patients than invasive collection techniques for detection of Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG). Self-collection is useful for Sexually Transmitted Infection...

2319 A TMPRSS2:ERG GENE FUSION MOLECULAR URINE ASSAY CORRELATES WITH PATHOLOGIC STAGE AND PROSTATECTOMY GLEASON SCORE AND IS ASSOCIATED WITH BIOPSY-TO-PROSTATECTOMY GLEASON UPGRADING

You have accessJournal of UrologyProstate Cancer: Markers1 Apr 20112319 A TMPRSS2:ERG GENE FUSION MOLECULAR URINE ASSAY CORRELATES WITH PATHOLOGIC STAGE AND PROSTATECTOMY GLEASON SCORE AND IS ASSOCIATED WITH BIOPSY-TO-PROSTATECTOMY GLEASON UPGRADING Alexander Haese, Felix Chun, Sarah Meyer, Sheila Aubin, John Day, and Jack Groskopf Alexander HaeseAlexander Haese Hamburg, Germany More articles by this author , Felix ChunFelix Chun Hamburg, Germany More articles by this author , Sarah MeyerSarah Meyer San Diego, CA More articles by this author , Sheila AubinSheila Aubin San Diego, CA More articles by this author , John DayJohn Day San Diego, CA More articles by this author , and Jack GroskopfJack Groskopf San Diego, CA More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2011.02.2566AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES An unmet need in PCa prognosis is the differentiation between indolent and significant PCa. Biochemical (e.g. PSA), clinical (e.g. clinical stage) or histological (BxGleason score) parameters, alone or combined, are not sufficiently accurate to aid in this critical distinction. On a genetic basis, a fusion between TMPRSS2 and the ETS family of transcription factors, found in 50–70% of PCa, is the most common specific gene rearrangement identified to date among solid human malignancies. Studies have linked PCa gene fusions to indicators of significant PCA. In this prospective, ongoing study, we evaluated the correlation of a prototype quantitative TMPRSS2:ERG (T2:ERG) gene fusion urine assay with outcomes in men with PCa scheduled for RRP. METHODS Post-DRE first-catch urine specimens were collected prior to RRP. T2:ERG mRNA copies were quantified using a transcription-mediated amplification assay and results normalized to PSA mRNA copies by calculating the ratio of TMPRSS2:ERG/PSA copies x 100,000 (T2:ERG Score). The prototype T2:ERG urine assay detects the gene fusion mRNA isoform corresponding to TMPRSS2 exon 1 to ERG exon 4. The T2:ERG Score was correlated with the presence or absence of pathologically organ (OC) vs. non-organ-confined (NOC) PCa and the presence or absence of Bx-to-RRP Gleason upgrading. RESULTS Of 74 subjects 28 (38%) had NOC-disease (≥pT3) and 69 (93%) had a RRP Gleason score ≥7. 21/26 subjects with BxGleason score of 6 were upgraded to Gleason Score ≥7 at RRP. Median T2:ERG Score was significantly higher in men with NOC vs. OC-PCa (80 vs. 9, p=0.002) and with RRP Gleason scores ≥7 vs. 6 (31 vs. 2, p=0.04). The median T2:ERG Score for subjects with vs without Bx-to-RRP-upgrading was 32 vs 2, respectively. CONCLUSIONS The T2:ERG assay correlated significantly with pT-stage and RRPGleason score. Interestingly, subjects with Bx-to-RRP-upgrading showed a trend towards higher median T2:ERG Scores. These data suggest that a T2:ERG urine assay may be useful to decide when more aggressive treatment of PCa is necessary. © 2011 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 185Issue 4SApril 2011Page: e930 Advertisement Copyright & Permissions© 2011 by American Urological Association Education and Research, Inc.MetricsAuthor Information Alexander Haese Hamburg, Germany More articles by this author Felix Chun Hamburg, Germany More articles by this author Sarah Meyer San Diego, CA More articles by this author Sheila Aubin San Diego, CA More articles by this author John Day San Diego, CA More articles by this author Jack Groskopf San Diego, CA More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

First Report of Performance of the Versant CT/GC DNA 1.0 Assay (kPCR) for Detection of Chlamydia trachomatis and Neisseria gonorrhoeae

We evaluated the analytical, work flow, and clinical performance of the Versant CT/GC DNA 1.0 assay (Versant CT/GC assay, where "CT" represents Chlamydia trachomatis and "GC" represents Neisseria gonorrhoeae). The assay simultaneously detects Chlamydia trachomatis and Neisseria gonorrhoeae in swab and first-catch urine (FCU) specimens. The limit of detection (LoD) was determined to be 342 copies/ml for C. trachomatis and 137 copies/ml for GC. The Versant CT/GC assay detected 15 C. trachomatis serovars and 46 GC strains. The Versant CT/GC assay demonstrated no cross-reactivity with 136 potentially cross-reacting organisms. Clinical concordance of the Versant CT/GC assay to the Aptima Combo 2 (AC2) assay from Gen-Probe was demonstrated using 1,129 patient specimens, including 589 urine and 540 swab specimens. Discrepant specimens were subjected to DNA sequencing to identify the presence of amplified targets and to identify false-positive and false-negative results. Overall percent agreement was greater than 98%. Positive and negative percent agreements for detection of C. trachomatis were 94.4% and 99.1%, respectively, in urine specimens and 95.8% and 99.8%, respectively, in swab specimens. Positive percent agreement for the detection of N. gonorrhoeae was 100% in both urine and swab specimens, and negative percent agreements were 99.6% and 99% in urine and swab specimens, respectively. In conclusion, the performance of the Versant CT/GC assay was comparable to that of the AC2 assay. The Versant CT/GC assay can be recommended for the detection of C. trachomatis and N. gonorrhoeae in swab and urine specimens of symptomatic and asymptomatic individuals.

Prevalence and Correlates of Trichomonas vaginalis Infection Among Female US Federal Prison Inmates

Previous studies have observed high prevalences of Trichomonas vaginalis infection among women entering US jails and state prisons (22%-47%). We sought to determine the prevalence among women incarcerated in 2 US female-only federal prisons. Female inmates were recruited at 2 prisons (n = 624). Participants completed a self-administered questionnaire and provided self-collected first-catch urine and vaginal swab specimens. Specimens were tested for T. vaginalis DNA. Approximately 8.5% of participants at the first prison, and 8.3% at the second prison had a positive urine result, vaginal swab result or both, for a combined prevalence of 8.5%. Using positivity in either specimen as the reference standard, urine polymerase chain reaction had a sensitivity of 66.7% and vaginal swab polymerase chain reaction had a sensitivity of 84.4%. The only significant positive correlate of T. vaginalis infection was lower household income before arrest. Other variables nonsignificantly positively correlated with T. vaginalis were being employed at the time of arrest, having experienced sexual, physical, or emotional abuse by a family member, having a parent who had not had a drug or alcohol addiction, never exchanging sex for money or drugs, ever being pregnant, having abnormal vaginal bleeding/spotting, and having concurrent chlamydia or gonorrhea. Although not as high as in other studies of women entering US jails and state prisons, our observed T. vaginalis prevalence of 8.5% was much higher than in the general US population. Therefore, screening for T. vaginalis infection may be warranted at federal prison entry, as well as sexual health education during prison stay.

1738 IMPROVED PREDICTION OF PROSTATE BIOPSY OUTCOME BY INCORPORATING TMPRSS2:ERG AND OTHER RISK FACTORS INTO A LOGISTIC REGRESSION MODEL

You have accessJournal of UrologyProstate Cancer: Detection and Screening I1 Apr 20101738 IMPROVED PREDICTION OF PROSTATE BIOPSY OUTCOME BY INCORPORATING TMPRSS2:ERG AND OTHER RISK FACTORS INTO A LOGISTIC REGRESSION MODEL James Amberson, John Day, James Cochran, William Harper, William Johnston, Naveen Kella, Isaac Koziol, George Patsias, and Thomas Williams James AmbersonJames Amberson Stratford, CT More articles by this author , John DayJohn Day San Diego, CA More articles by this author , James CochranJames Cochran Dallas, TX More articles by this author , William HarperWilliam Harper Columbus, GA More articles by this author , William JohnstonWilliam Johnston Hyannis, MA More articles by this author , Naveen KellaNaveen Kella San Antonio, TX More articles by this author , Isaac KoziolIsaac Koziol Richmond, VA More articles by this author , George PatsiasGeorge Patsias Wellington, FL More articles by this author , and Thomas WilliamsThomas Williams Sarasota, FL More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2010.02.1586AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES The need exists for prostate cancer (PCa) markers with greater diagnostic accuracy. Although serum prostate specific antigen (sPSA) has high sensitivity for predicting prostate biopsy (PB) outcome, it has poor specificity leading to >60% negative biopsies. In this study, we evaluated the performance of a prototype quantitative TMPRSS2:ERG (T2:ERG) gene fusion urine assay for predicting PB outcome alone and in combination with other markers and risk factors. METHODS Patients referred for PB because of abnormal DRE or elevated sPSA were enrolled in a prospective clinical study at 7 community-based urology clinics located throughout the United States. Post-DRE first-catch urine specimens were collected from 471 subjects, who then had PB performed (minimum 12 cores). T2:ERG mRNA copies in samples were quantified using a transcription-mediated amplification assay and normalized to PSA mRNA copies. The prototype T2:ERG urine assay detects the gene fusion mRNA isoform TMPRSS2 exon 1 to ERG exon 4. Logistic regression (LR) analysis was used to determine the best combination of risk factors and biomarkers for predicting PB outcome. RESULTS In this study, 98% (463/471) of specimens yielded sufficient RNA for T2:ERG analysis. PCA3, sPSA and fPSA results were obtained for 453 subjects, and the positive PB rate was 44% (199/453). The T2:ERG urine assay showed high specificity (87%) and a sensitivity of 39% using a cutoff of 32. Combining T2:ERG with PCA3, sPSA, %fPSA, age, prostate volume, family history, race, DRE result and prior prostate biopsy in a cross-validated LR analysis led to significantly improved diagnostic accuracy (AUC=0.754, 95% CI=0.749-0.758) for predicting PB outcome. The complete model without T2:ERG had an AUC of 0.715 (95% CI=0.710-0.720). CONCLUSIONS In a prospective multi-center study, the T2:ERG urine assay demonstrated high specificity for predicting biopsy outcome; sensitivity was similar to the prevalence of T2:ERG gene fusions in prostate tumors. These data suggest that further improvement in diagnostic accuracy could be achieved using a nomogram that combines T2:ERG with other markers and risk factors for prostate cancer. © 2010 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 183Issue 4SApril 2010Page: e671-e672 Advertisement Copyright & Permissions© 2010 by American Urological Association Education and Research, Inc.MetricsAuthor Information James Amberson Stratford, CT More articles by this author John Day San Diego, CA More articles by this author James Cochran Dallas, TX More articles by this author William Harper Columbus, GA More articles by this author William Johnston Hyannis, MA More articles by this author Naveen Kella San Antonio, TX More articles by this author Isaac Koziol Richmond, VA More articles by this author George Patsias Wellington, FL More articles by this author Thomas Williams Sarasota, FL More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

The Diaphragm and Lubricant Gel for Prevention of Cervical Sexually Transmitted Infections: Results of a Randomized Controlled Trial

BackgroundWe evaluated the effectiveness of the Ortho All-Flex Diaphragm, lubricant gel (Replens®) and condoms compared to condoms alone on the incidence of chlamydial and gonococcal infections in an open-label randomized controlled trial among women at risk of HIV/STI infections.MethodsWe randomized 5045 sexually-active women at three sites in Southern Africa. Participants who tested positive for curable STIs were treated prior to enrollment as per local guidelines. Women were followed quarterly and tested for Chlamydia trachomatis (CT) or Neisseria gonorrhoeae (GC) infection by nucleic-acid amplification testing (Roche Amplicor®) using first-catch urine specimens. STIs detected at follow-up visits were treated. We compared the incidence of first infection after randomization between study arms in both intent-to-treat (ITT) and per-protocol populations.FindingsBaseline demographic, behavioral and clinical characteristics were balanced across study arms. Nearly 80% of participants were under 35 years of age. Median follow-up time was 21 months and the retention rate was over 93%. There were 471 first chlamydia infections, 247 in the intervention arm and 224 in the control arm with an overall incidence of 6.2/100 woman-years (wy) (relative hazard (RH) 1.11, 95% Confidence Interval (CI): 0.93–1.33; p = 0.25) and 192 first gonococcal infections, 95 in the intervention arm and 97 in the control arm with an overall incidence of 2.4/100wy (RH 0.98, 95%CI: 0.74–1.30; p = 0.90). Per protocol results indicated that when diaphragm adherence was defined as “always use” since the last visit, there was a significant reduction in the incidence of GC infection among women randomized to the intervention arm (RH 0.61, 95%CI: 0.41–0.91, P = 0.02).InterpretationThere was no difference by study arm in the rate of acquisition of CT or GC. However, our per-protocol results suggest that consistent use of the diaphragm may reduce acquisition of GC.Trial RegistrationClinicalTrials.gov NCT00121459 [NCT00121459]

Epidemiological, social, diagnostic and economic evaluation of population screening for genital chlamydial infection

To investigate epidemiological, social, diagnostic and economic aspects of chlamydia screening in non-genitourinary medicine settings. Linked studies around a cross-sectional population-based survey of adult men and women invited to collect urine and (for women) vulvovaginal swab specimens at home and mail these to a laboratory for testing for Chlamydia trachomatis. Specimens were used in laboratory evaluations of an amplified enzyme immunoassay (PCE EIA) and two nucleic acid amplification tests [Cobas polymerase chain reaction (PCR), Becton Dickinson strand displacement amplification (SDA)]. Chlamydia-positive cases and two negative controls completed a risk factor questionnaire. Chlamydia-positive cases were invited into a randomised controlled trial of partner notification strategies. Samples of individuals testing negative completed psychological questionnaires before and after screening. In-depth interviews were conducted at all stages of screening. Chlamydia transmission and cost-effectiveness of screening were investigated in a transmission dynamic model. General population in the Bristol and Birmingham areas of England. In total, 19,773 women and men aged 16-39 years were randomly selected from 27 general practice lists. Screening invitations reached 73% (14,382/19,773). Uptake (4731 participants), weighted for sampling, was 39.5% (95% CI 37.7, 40.8%) in women and 29.5% (95% CI 28.0, 31.0%) in men aged 16-39 years. Chlamydia prevalence (219 positive results) in 16-24 year olds was 6.2% (95% CI 4.9, 7.8%) in women and 5.3% (95% CI 4.4, 6.3%) in men. The case-control study did not identify any additional factors that would help target screening. Screening did not adversely affect anxiety, depression or self-esteem. Participants welcomed the convenience and privacy of home-sampling. The relative sensitivity of PCR on male urine specimens was 100% (95% CI 89.1, 100%). The combined relative sensitivities of PCR and SDA using female urine and vulvovaginal swabs were 91.8% (86.1, 95.7, 134/146) and 97.3% (93.1, 99.2%, 142/146). A total of 140 people (74% of eligible) participated in the randomised trial. Compared with referral to a genitourinary medicine clinic, partner notification by practice nurses resulted in 12.4% (95% CI -3.7, 28.6%) more patients with at least one partner treated and 22.0% (95% CI 6.1, 37.8%) more patients with all partners treated. The health service and patients costs (2005 prices) of home-based postal chlamydia screening were 21.47 pounds (95% CI 19.91 pounds, 25.99) per screening invitation and 28.56 pounds (95% CI 22.10 pounds, 30.43) per accepted offer. Preliminary modelling found an incremental cost-effectiveness ratio (2003 prices) comparing screening men and women annually to no screening in the base case of 27,000 pounds/major outcome averted at 8 years. If estimated screening uptake and pelvic inflammatory disease incidence were increased, the cost-effectiveness ratio fell to 3700 pounds/major outcome averted. Proactive screening for chlamydia in women and men using home-collected specimens was feasible and acceptable. Chlamydia prevalence rates in men and women in the general population are similar. Nucleic acid amplification tests can be used on first-catch urine specimens and vulvovaginal swabs. The administrative costs of proactive screening were similar to those for opportunistic screening. Using empirical estimates of screening uptake and incidence of complications, screening was not cost-effective.

Vulvovaginal-Swab or First-Catch Urine Specimen To Detect Chlamydia trachomatis in Women in a Community Setting?

Screening for chlamydia in women is widely recommended. We evaluated the performance of two nucleic acid amplification tests for detecting Chlamydia trachomatis in self-collected vulvovaginal-swab and first-catch urine specimens from women in a community setting and a strategy for optimizing the sensitivity of an amplified enzyme immunoassay on vulvovaginal-swab specimens. We tested 2,745 paired vulvovaginal-swab and urine specimens by PCR (Roche Cobas) or strand displacement amplification (SDA; Becton Dickinson). There were 146 women infected with chlamydia. The assays detected 97.3% (95% confidence interval [CI], 93.1 to 99.2%) of infected patients with vulvovaginal-swab specimens and 91.8% (86.1 to 95.7%) with urine specimens. We tested 2,749 vulvovaginal-swab specimens with both a nucleic acid amplification test and a polymer conjugate-enhanced enzyme immunoassay with negative-gray-zone testing. The relative sensitivities obtained after retesting specimens in the negative gray zone were 74.3% (95% CI, 62.8 to 83.8%) with PCR and 58.3% (95% CI, 46.1 to 69.8%) with SDA. In community settings, both vulvovaginal-swab and first-catch urine specimens from women are suitable substrates for nucleic acid amplification tests, but enzyme immunoassays, even after negative-gray-zone testing, should not be used in screening programs.

Quantifying leukocytes in first catch urine provides new insights into our understanding of symptomatic and asymptomatic urethritis

We quantitatively investigated inflammatory cells in the male urethra. Leukocytes in the first catch urine (FCU) from 87 men with and without urethritis were quantitated using haemocytometer counts and stained with an anti-CD45 pan-leukocyte antibody. An increased number of leukocytes in FCU specimens was associated with urethritis (P > 0.002), the presence of discharge and/or dysuria (P < 0.001), and detection of Chlamydia trachomatis (P < 0.001) and Neisseria gonorrhoeae (P < 0.001). In men with urethritis, higher leukocyte counts were also observed in the above groups (P = 0.07, 0.03 and P < 0.0001, respectively). As leukocyte number increased, the likelihood of detecting either pathogen increased. This study suggests that symptoms and signs are a surrogate marker for the degree of inflammation present, and that as urethral inflammation increases, the likelihood of detecting a sexually transmitted pathogen also increases. This would explain why men with asymptomatic urethritis are less likely to have a sexually transmitted infection detected than those with discharge and/or dysuria.