bluuebunny/crossref_metadata_2025 huggingface dataset:
P1-S1.41 Emergence of an African antimicrobial resistance genotype inNeisseria gonorrhoeaestrains isolated in Guangzhou, China, 2001–2009
Authors: Zheng H P , Yang B , Wu X Z , Huang J M , Zeng W Y , Xue Y H , Jiang H N , Ou J L
Published: July 2011

Background

The continuing spread of drug-resistant gonococci has posed a challenge for successful treatment worldwide. Recently third-generation cephalosporins-resistant strains were isolated in Japan. Guangdong Province in South China has one of the highest gonococcal resistance rates in China and a large number of international migrants. We investigated the invitro antimicrobial susceptibility and genotypes ofN gonorrhoeaestrains isolated in Guangzhou, the capital city, from 2001 to 2009.

Methods

MICs to penicillin, ceftriaxone, tetracycline, ciprofloxacin, and spectinomycin were determined by agar plate dilution and susceptibilities were interpreted according to WHO standards. ß-lactamase production was determined by paper acidometric testing. The resistant plasmids were determined for penicillinase-producingN gonorrhoeae(PPNG) and high-level tetracycline resistantN gonorrhoeae(TRNG) by PCR and the isolates were genotyped.

Results

Of 1132 consecutive gonococci isolated from 2001 to 2009, no ceftriaxone and spectinomycin resistant strains were found, but the prevalence of strains less susceptible to ceftriaxone rose from 18.2% to 38.5%. The MIC90 for ceftriaxone showed intermediate sensitivity (0.06–0.125 μg/ml) and spectinomycin near the resistant level (16–32 μg/ml). The resistance to penicillin, tetracycline and ciprofloxacin increased from 81.8%, 84.5% and 72.5% in 2001 to 90.1%, 91.4% and 98.7% in 2009, respectively. A total of 313 (27.7%) PPNG and 486 (42.9%) TRNG strains were detected. 206 (18.2%) strains were both PPNG and TRNG. PPNG rose from 21.8% to 29.8% and TRNG rapidly increased from 27.2% to 56.3%. Genotyping TEM-1 gene showed 267 (99.6%) PPNGs in 2001–2008 carried the Asian-type ß-lactamase plasmids and one African-type in 2008. Genotyping of tetM gene showed that all 486 TRNGs were Dutch variants.

Conclusion

Gonorrhoea resistance continues to be a major public health problem in Guangzhou. The emergence of an African gonorrhoea resistance variant may be related to the large African diaspora in Guangzhou, migration of Chinese to Africa, or other migration patterns. More research is needed to determine what practices, systems, and behaviours contribute to escalating resistance patterns.

Abstract P1-S1.41 Figure 1

Resistance ofN gonorrhoeaeisolates in Guangzhou during 2001–2009.
P3-S1.13 Establishment of a protocol for the detection ofChlamydia trachomatisin semen specimens using the cobas®4800 CT/NG test
Authors: de Barbeyrac B , Mehats V , Papaxanthos A , Bébéar C
Published: July 2011

Background

In addition to common urogenital samples, semen specimens are used to detectChlamydia trachomatis(CT). In France, testing of semen specimens is mandatory by law in context of donors and in vitro fertilisation. However, semen specimens are known to show significant inhibition in PCR assays. Analysis of cycles of threshold (ct) values can be used to determine inhibition and limits of detection of assays.

Objective

To establish a suitable protocol for the highly sensitive detection of CT in semen specimens using the fully automated cobas 4800® CT test.

Materials and Methods

We evaluated inhibition rates and limits of detection (LOD) for different semen sample volumes by ct value analysis in the cobas® 4800 CT test. Semen samples were obtained from 100 patients visiting the reproduction biology department of the Bordeaux University hospital. Between 5 and 67 replicates of semen ranging from 1 to 50 μl were each added to 4.5 ml of cobas® PCR media and run in the cobas® 4800 CT test. The inhibition rate was compared to the inhibition rate obtained in 323 urogenital swabs and 278 urines using the ct values of the internal control (IC). Thereafter, semen volumes that showed the lowest inhibition rates were selected to determine the LOD, by comparing mean ct values for the target in the cobas® 4800 CT test using semen specimens spiked with different concentrations of CT.

Results

Mean IC ct values in semen volumes ranging from 1 to 40 μl did not differ from those obtained in urogenital swabs and urines, whereas semen volumes of 50 μl resulted in a marked increase in IC ct values indicating inhibition. Therefore, semen volumes of 25, 40, and 50 μl diluted in 4.5 ml of cobas® PCR media were tested to determine the LOD. Mean ct values generated by the target CT are shown in Abstract P3-S1.13 table 1. Ct values were similar in samples of 25 μl semen and controls, but were higher or even negative in 40 and 50 μl semen volumes indicating that a semen volume of 25 μl showed the lowest LOD (10–5) combined with a low inhibition rate.

Abstract P3-S1.13 Table 1

Comparison of ct values in different semen sample volumes spiked with serial dilutions of CT-infected cells

Ct valuesSerial dilutions of CT-infected cellsControl sample25 μl semen40 μl semen50 μl semen10-13130.230.632.610-233.233.332.835.810-335.736.835.337.510-437.937.13840.510-54040.0negneg

Conclusion

Semen specimens can be tested in the cobas® 4800 CT test applying our easy to perform, highly sensitive, and low inhibition protocol.
P3-S1.16 Comparison of the Abbott m2000 RealTime CT assay forChlamydia trachomatismonitoring in Tanzania compared to the Roche Amplicor CT assay
Authors: Agreda P , Dize L , West S , Williams J A , Van Der Pol B , Gaydos C A
Published: July 2011

Background

The Abbott m2000 RealTime CT assay was evaluated as a new option for the detection ofChlamydia trachomatisin specimens obtained from Tanzania, and its performance was compared to the performance of the Roche Amplicor CT Assay.

Methods

Duplicate swab specimens were collected from villages throughout Tanzania. 304 duplicate samples were shipped to the Johns Hopkins STD laboratory at 2–8°C for the detection ofC trachomatis. All swab specimens were shipped in a dry state, expressed in 1 ml of sterile molecular grade DEPC water upon arrival, and analysed using the Roche Amplicor CT assay. Prior to Roche Amplicor amplification and detection, DNA extraction was performed using the Roche Magna Pure LC robot. The duplicate swab specimens were shipped to Indiana University for Abbott m2000 RealTime CT assay analysis. The bacterial load was measured by the DC value of the m2000 RealTime CT.

Results

Of 304 specimens, 44 (14.5%) were positive for CT via Roche Amplicor CT assay, and 53 (17.4%) were positive for CT via Abbott m2000 RealTime CT assay. The relative quantitation for the m2000 assay ranged from DC 0.62 to DC 22.16. If the Roche PCR assay was considered to be the reference standard, the Abbott m2000 RealTime CT assay sensitivity was 44/44 (100%), specificity was 251/260 (96.53%), positive predictive value was 251/251(100%), and negative predictive value was 44/53 (83.01%). The κ score was 0.890. Discordant specimens, which were determined to be negative by Roche Amplicor and positive by Abbott m2000 RealTime, were tested by Gen-Probe TMA. Of nine discordant tests, two were positive, five were negative, and two were of insufficient volume for retesting. After discordant testing, there appeared to be five samples that were graded to be false positives by m2000. The Abbott m2000 RealTime CT assay sensitivity remained 100%, while specificity increased to 256/258 (99.2%). The negative predictive value increased to 46/48 (95.83%). The κ score was 0.9748 after discordant results were further analysed.

Conclusions

Abbott m2000 RealTime CT assay demonstrates excellent sensitivity and specificity compared to the Roche Amplicor CT Assay for the detection ofC trachomatis. It may be advantageous to be able to measure the relative concentration for CT in some epidemiologic studies.
P3-S1.17 Syndromic management of cervicitis and vaginal discharge at a STI clinic in Jamaica: low cure rates for Chlamydial infection and trichomoniasis
Authors: Hobbs M , Anderson C , Hylton-Kong T , Eastman S , Rich K , Gallo M , Legardy-Williams J , Costenbader E , Steiner M , Warner L
Published: July 2011

Background

Management of cervicitis and abnormal vaginal discharge in Jamaica is based on the syndromic approach recommended by the WHO. To evaluate current algorithms for treatment of gonorrhoea, chlamydial infection and trichomoniasis, we conducted laboratory testing with vaginal specimens from women presenting with cervicitis or vaginitis syndromes at a sexually transmitted infections clinic in Kingston and at follow-up to assess cure rates for these infections.

Methods

From August, 2010 through January, 2011, vaginal swab specimens were obtained from 258 women >18 years old during a routine clinical examination prior to syndromic treatment according to local guidelines. Treatment for gonorrhoea, chlamydial infection and trichomoniasis was prescribed for women with cervicitis. Treatment for trichomoniasis, bacterial vaginosis and candidiasis was prescribed for women with abnormal vaginal discharge. Women returned the next week for follow-up assessment and specimen collection. Specimens were tested forNeisseria gonorrhoeae(NG) andChlamydia trachomatis(CT) using APTIMA Combo2 and forTrichomonas vaginalis(TV) using APTIMA analyte-specific reagents. All baseline specimens were tested. Follow-up specimens from women with a positive baseline test or who remained symptomatic were also tested. Patients with a positive follow-up test were contacted and instructed to return to the clinic for additional treatment. Cure was defined as a positive baseline test and a negative follow-up test.

Results

Baseline prevalence of infection with NG was 11.7%, CT was 20.7%, TV was 25.6%. At least one of these STIs was detected by laboratory testing in 40.7% of women. Co-infections were common. Women with TV were more likely to have NG or CT than women without TV (OR: 2.6, 95% CI 1.4 to 4.8). STI testing at follow-up indicated cure rates of 77.3% for NG, 43.5% for CT and 47.1% for TV infections. CT incidence at follow-up was 5.9%; no incident NG or TV infections were detected.

Conclusions

With syndromic management, just over half of the STIs in women that were detected by laboratory testing at baseline were cured at follow-up. Reinfection, incorrect or inadequate treatment, failure to comply with treatment instructions or treatment failure could potentially explain prevalent STIs that were detected at follow-up. The low cure rates for chlamydial infection and trichomoniasis are cause for concern.
P3-S1.18 Side effects of doxycycline in adolescents treated for pelvic inflammatory disease
Authors: Risser W , Risser J
Published: July 2011

Background

Our purpose was to assess adverse reactions to doxycline in adolescents under treatment for PID.

Methods

At the Harris County (Houston), Texas, USA Juvenile Detention Center from May 2008 through January 2010, we evaluated adolescents in whom we diagnosed PID. We used the diagnostic criteria and outpatient antibiotic regimen for PID recommended by the CDC. We gave 250 mg ceftriaxone intramuscularly once, and, if they were not pregnant, vomiting, or ill enough to also require metronidazole, we prescribed doxycline, 100 mg twice a day for 14 days. The doxycycline was given before, during, or after breakfast, and approximately 2 h after supper. With the medicine, patients were routinely given a packet of two crackers. We re-assessed these patients at 48–72 h and again at least at 7 and 14 days. At each follow-up, we discussed side effects of doxycycline.

Results

We evaluated 141 consecutive patients in whom we made the diagnosis of mild to moderate PID: 55% were black, 31% Hispanic, and 15% white. The mean age (SD) was 15.4 (1.2) years. Of the 141 patients, 86 (61%) had no problems when the doxycline was given with two crackers. Forty-five (32%) required more substantial food to avoid nausea and/or vomiting: the medication was given immediately after breakfast or with a sandwich after supper. Those who had side effects in the morning had usually skipped breakfast. Ten (7%) complained of significant gastrointestinal side effects that were not relieved by the simultaneous ingestion of food, and we gave a dose of promethezine 1 h before the doxycycline.

Conclusion

39% of our patients required more than a small amount of food (two crackers) to avoid gastrointestinal side effects from doxycline. Our results suggest that clinicians should advise adolescents to take this antibiotic after eating a greater amount of food if a small snack does not prevent this problem. Because some patients (7%) did not tolerate doxycline even after they ate a meal or a sandwich, we also recommend early contact with patients who have PID to assess the need to change medications or to add an anti-emetic.
P3-S1.23 Mycoplasma genitaliumin South African men with and without symptoms of urethritis: diagnosis and bacterial load
Authors: Hoosen A , le Roux M C , Adam A
Published: July 2011

Objectives

To diagnose causes of urethritis in symptomatic men and evaluate the role ofMycoplasma genitaliumby calculating bacterial loads in urine specimens from symptomatic and asymptomatic men.

Methods

Urine specimens from 94 men with visible urethral discharge, 206 with burning on micturition (BOM) and 75 without any genital symptoms were tested forN gonorrhoeae(NG),C trachomatis (CT), T vaginalis (TV) and M genitalium(MG) using TMA assays. The bacterial load was estimated by quantitative real-time PCR assay in specimens withM genitaliumalone.

Results

At least one pathogen was detected in 138 (46%); mixed infections in 28 (9.3%). The most common mixed infection was NG + MG (3.7%). There were significant differences for MG (17.3% vs 6.6%), NG (16.7% vs 2.6%), CT (12.3% vs 5.1%), and TV (8.0% vs 1.3%) in symptomatic vs asymptomatic men. There was also a significant difference in presence of MG in the symptomatic men (20.2% in men with discharge vs 4.4% in those with BOM). Patients with urethral discharge (mean 5.3×103 copies/ul) had significantly higherM genitaliumconcentrations than those with BOM (mean 6.7×102 copies/ul) (p<0.001), and the bacterial concentrations in men with symptoms and/or signs of urethritis were significantly higher than that in asymptomatic men (mean 4.6×101 copies/ul) (p=0.02).

Conclusions

Significantly more organisms were detected in patients with urethritis than in those without. There was a strong association betweenM genitaliumbacterial load and men with symptoms of urethritis both BOM and urethral discharge. As the number of organisms increased, the severity of the symptoms increased (greater numbers in urethral discharge), indicating a role forM genitaliumin disease severity. This study also demonstrated the usefulness of quantitative PCR in determining an aetiological role and possibility of monitoring efficacy of treatment for this pathogen.
P3-S1.24 Cost effectiveness of self-taken vaginal swabs using GenProbe AC2 assay vs clinician taken urethral and endocervical swabs for detection of gonorrhoea
Authors: Wilson J , Hulme C , Stewart C , Schoeman S , Wilcox M
Published: July 2011

Background

Culture ofNeisseria gonorrhoeaefrom the urethral and endocervix is currently the recommended method of detecting gonorrhoea (GC) in women in the UK. This necessitates a speculum examination. NAATs are the mainstay of testing forChlamydia trachomatis(CT) infection and assays exist which can also detect GC. NAATs are more sensitive than culture meaning non-invasive samples can be used. No previous study has compared self-taken VVS analysed by AC2 with urethral and endocervical GC culture, which is considered the gold standard test in the UK. We therefore undertook this comparison in a clinical setting with low GC prevalence and performed a cost-effectiveness analysis.

Methods

Participants were recruited within the Centre for Sexual Health at Leeds, UK between March 2009 and January 2010. The women, presenting for a new visit, performed a self-taken VVS and were then examined and urethral and endocervical samples taken for GC culture. Positive AC2 tests were confirmed by the Aptima GC and/or Aptima CT mono-specific platform tests. The cost effectiveness was analysed using incremental cost effectiveness ratios (ICERs). The analysis uses correct detection of GC as the primary endpoint and adopts the perspective of the UK National Health Service. Costs were estimated from a sub-sample of 92 patients in July 2009. The analysis includes costs of clinic time, together with the laboratory costs. Comparisons were made of self-taken VVSs using AC2 vs clinician taken cultures for the whole cohort.

Results

100 of the 3973 participants (2.5%) were infected with GC. The overall sensitivity of GC culture was 82% and self-taken VVS 95%. 35% of the participants were asymptomatic in whom GC culture sensitivity was 79% and self-taken VVS 91%. The cost effectiveness results are presented in Abstract P3-S1.24 table 1.

Abstract P3-S1.24 Table 1

Diagnostic test

Diagnostic testCostsDifferenceGonorrhoea cases correctly diagnosedDifferenceICERs (£ per correctly diagnosed case)All have GC culture test and CT NAAT irrespective of symptoms250 841.7929 922.498213GC VVS dominatesAll have GC/CT VVS irrespective of symptoms220 919.3095

Conclusions

Self-taken VVS analysed by AC2 are cost effective compared to urethral and endocervical GC culture; being more sensitive and less costly. Whilst those presenting with symptoms continue to require a clinician examination use of VVSs in women with no symptoms removes the need for examination. The cost of the time taken in explanation of self taken swabs for this group represents <20% of the cost of a clinician examination. Cost savings also accrue through use of AC2 alone rather than GC culture and CT NAAT.
P3-S1.25 Evaluation of real time PCR for detection of five sexually transmitted diseases in Abidjan, Ivory Coast
Authors: Kakou-Ngazoa S
Published: July 2011

Objective

To evaluate a real-time PCR assay for molecular screening of five most common sexually transmitted diseasesTrichomonas vaginalis, Mycoplasma genitalium, Neisseria gonorrhoeae, Chlamydia trachomatisand Herpes simplex virus 2 in Abidjan, Ivory Coast.

Methods

25 self-collected vaginal swabs were obtained from adult persons with symptoms of vaginal discharge and vulval irritation. The samples were diluted in sterile PBS and the DNA extraction was processed using Nuclisens extraction Kit. 5 μl of genome extract was applied in real-time PCR for each pathogen. The targeted region of real time was the 18 S ribosomal DNA genes forT vaginalisand the 16S rRNA forM genitalium.

Results

10 of 25 (40%) samples were positive for sexually transmitted dieases. Over 30 % of the samples shown inhibition and were tested positive by serial dilution.Chlamydia trachomatisis the most detected infection in five samples, and one sample was positive respectively byT vaginalisand byN gonorrhoeae. We detect no co-infection in all samples.

Conclusions

The real-time PCR is a suitable specific and sensitive method to investigate and to detect sexually transmitted diseases in developing countries.
P3-S1.28 Is urethritis of unknown aetiology caused by bacteria associated with bacterial vaginosis?
Authors: Frølund M , Datcu R , Ahrens P , Lidbrink P , Björnelius E , Jensen J S
Published: July 2011

Background

The aetiology of non-gonococcal urethritis (NGU) in men is unknown in 30%–50% of cases. Little is known about the relation of bacterial vaginosis (BV) associated bacteria in men with urethritis of unknown aetiology (UUE). Urethral swabs from men with and without NGU were analysed with PCR for BV associated bacteria to show a possible association with UUE.

Methods

Urethral swabs from 9 and 19 men with symptomatic and asymptomatic NGU (>5 PMNL/hpf), respectively, and 30 asymptomatic men without NGU were collected. All samples were negative forNeisseria gonorrhoeae, Chlamydia trachomatis, Mycoplasma genitalium,Ureaplasma urealyticumandU parvumwith specific PCR assays. Quantitative real-time PCR with TaqMan based assays were performed to detect Atopobium vaginae (Av), Sneathia sanguinegens (Ss), Leptotrichia amnionii (La) and Gardnerella vaginalis (Gv), and with SYBR green assays for BVAB 1, 2 and 3, and Megasphaera phylotype 1 (M1).

Results

Gv was detected in 93% of cases with UUE and in 37% of controls (p<0.0001). There was no difference in organism load. In the 28 NGU cases Av, Ss and La were found in 3, 2 and 1 samples, respectively, and in 6, 2 and 1 of the control samples, respectively. The median corresponding organism loads were 14, 95 and 51 for the NGU cases and 16, 10 566 and 353 for the controls. All samples were negative for BVAB 1, 2 and 3 and M1, except one control with 10 genome copies of BVAB 1.

Conclusions

Gardnerella vaginaliswas associated with male urethritis in this study, especially in men with asymptomatic urethritis, while BVAB 1, 2 and 3, Megasphaera phylotype 1,Atopobium vaginae,Sneathia sanguinegens, andLeptotrichia amnioniiwere not.
P1-S1.43 Antimicrobial susceptibility and molecular epidemiological characteristics ofNeisseria gonorrhoeaein 2009–2010 in Belarus
Authors: Glazkova S , Unemo M , Golparian D , Titov L , Pankratova N , Suhabokava N , Shimanskaya I , Ignatyev G , Domeika M
Published: July 2011

Background

Antimicrobial resistance (AMR) inNeisseria gonorrhoeaeis a global concern, and gonorrhoea may become untreatable. However, AMR data from Eastern Europe are scarce beyond Russia, and in Belarus no AMR data or other characteristics of gonococci have been published in over 20 years. The aim was to describe the AMR and molecular epidemiological characteristics of gonococci circulating in 2009 and 2010 in Belarus.

Methods

N gonorrhoeaeisolates cultured in 2009 (n=80) and 2010 (n=78) in the two different cities Minsk (n=45) and Mogilev (n=113) were examined using Etest for nine antimicrobials and nitrocefin solution for β-lactamase production. Where available, breakpoints for susceptibility and resistance according to The European Committee on Antimicrobial Susceptibility Testing (EUCAST;http://www.eucast.org) were used. Moreover, screening for penA mosaic alleles, full-length porB gene sequencing, andN gonorrhoeaemultiantigen sequence typing (NG-MAST) were performed.

Results

The levels of resistance in 2009–2010 to antimicrobials evidently used in the gonorrhoea treatment in Belarus were—ceftriaxone 0%, spectinomycin 0%, azithromycin 14.4%, tetracycline 30.9%, ciprofloxacin 34.6%, and erythromycin 59.2% (only tested in 2009). The levels of resistance to other antimicrobials of international interest but not used in Belarus were—cefixime 0%, gentamicin 1.3%, and penicillin G 9.9% (only tested in 2009). Extraordinarily, no β-lactamase producing isolates were detected. The circulatingN gonorrhoeaepopulation identified was very heterogeneous and contained many divergent NG-MAST STs, of which more than half have not been previously described worldwide.

Conclusions

Due to the high levels of resistance to all antimicrobials previously recommended as first-line treatment, only ceftriaxone and spectinomycin can be recommended for empirical gonorrhoea treatment in Belarus. Continuous and quality assured gonococcal AMR surveillance in Eastern Europe is crucial, in Belarus this surveillance is now initiated using WHO protocols.
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
Authors: Ferrero D V , Biglino A
Published: July 2011

Background

IncreasingChlamydia trachomatis(CT) rates and ever-presentNeisseria gonorrhoeae(NG) infections worldwide in women have led to the consideration of targeted male CT/NG screening programs in order to address the male reservoir. Effortlessly collected, non-invasive, painless, dry, self-obtained male swab specimens that are stable and easy to transport would enhance the success of male screening programs. We undertook a study to compare the results of self obtained glans/meatal swab (SOGS) (COPAN FLOQSwabs) specimens with conventional first catch urine (FCU) specimens. The aim of the study was simply to assess the usefulness of a SOGS specimen when compared to FCU given the SOGS specimen's ease of collection, ease of use and handling in challenged venues (ie, without toilet facilities), and ease of transport.

Methods

122 male patients attending an STD clinic and males incarcerated near the Italian city of Asti were tested for CT and NG using a SOGS. Patients were verbally instructed and shown a drawing to enhance standardised self-collection. A FCU specimen was also collected. SOGS and FCU specimens were then processed according to the manufacturer's recommended procedure using the BD ProbeTec ET system (Becton Dickinson and Company).

Results

There were a total of 16 specimens detected positive for either CT or NG. Seven men were infected by both CT and NG. The CT prevalence was 5.7%. The NG prevalence was 13.1%. There was an overall SOGS specimen agreement of 97.5% when compared to FCU specimen results.

Conclusions

High agreement was obtained with the self obtained glans/meatal COPAN FLOQSwabs when compared to standard FCU specimens. These results indicate that self obtained glans/meatal swab specimens are a viable specimen choice. Dry self obtained glans/meatal swab specimens with FLOQSwabs were easy to collect, transport and test using the BD ProbeTec ET system. The BD ProbeTec ET system showed high agreement when testing both of these specimen types. Simpler methods of collection and transport that produce reliable test results may increase the potential for screening difficult to test male populations, especially those who never test.
P3-S1.32 A validation study of the Gen-Probe APTIMA Combo2 (AC2) assay for detectingChlamydia trachomatisandNeisseria gonorrhoeaein dry swabs
Authors: Cosentino L , Schwebke J R , Hobbs M M , Hillier S L
Published: July 2011

Background

Vaginal swabs are an optimal specimen for detection ofChlamydia trachomatis(CT) andNeisseria gonorrhoeae(GC). The Gen-Probe Aptima system (Aptima) requires use of a sample transport media, whereas the Becton Dickinson ProbeTec ET System (Probetec) can utilise dry swabs. The use of dry swabs allows for collection of STD test samples at non-clinical testing venues.

Methods

Swabs were collected from 180 sexually active women aged 15–25 years, who had asymptomatic BV and two or more risk factors for STDs. The participants received home vaginal swab self-collection kits for BV and STD testing. Participants mailed the kits directly to the lab. Probetec swabs were tested within 14 days of collection. A dry transport swab was placed into an Aptima vaginal swab collection tube, generally within 14 days of collection and stored at −80°C. Aptima swabs were thawed and tested in batches. Specimens with discordant results in the two nucleic acid amplification test systems were retested with both systems.

Results

There were 58 women (32%) positive for GC and 62 (34%) positive for CT. The level of agreement between the Aptima and Probetec systems was higher for CT (176/180, 98%) than for GC (171/180, 95%). Of the 13 samples with discordant results, five were resolved with repeat testing. All eight remaining samples had discordant GC results: seven were Probetec positive, Aptima negative and one was Probetec negative, Aptima positive.

Conclusions

Vaginal swabs tested in the Aptima system were equivalent to Probetec in detecting CT but were less sensitive for the detection of GC. Dry swabs cannot be recommended for detection of GC from vaginal swabs using the Aptima system.
P3-S1.33 Evaluation of the Roche cobas 4800 for the detection ofChlamydia trachomatisandNeisseria gonorrhoeaeusing minimally invasive samples in women
Authors: Hires L , Van Der Pol B , Taylor S , Lillis R , Body B
Published: July 2011

Background

The Roche cobas 4800 (cobas) is a new diagnostic assay that utilises an automated workstation to isolate nucleic acids from clinical specimens and a real time instrument for the detection ofC trachomatis(CT) andN gonorrhoeae(NG). The objective of this study was to compare the performance characteristics of the cobas to the BD Viper (Viper) and GenProbe Aptima Combo 2 (AC2) assays for the detection of CT and NG using a patient infected standard (PIS).

Methods

Specimens were obtained from women attending STD, family planning, or OB/GYN clinics from 12 geographically distinct locations. Urine and vaginal swabs were obtained from each participant as were endocervical (data not shown) and liquid based cytology samples (data not shown). Women were randomised to either self-obtained (SOV) or clinician-obtained (COV) vaginal swab collection. Four sites performed testing by AC2, cobas, and Viper for urine and by cobas for vaginal swabs. A patient was considered infected if at least two of the assays with different molecular targets gave positive results from cervical or urine samples.

Results

Overall CT sensitivity ranged from 91.9% to 93.9% and specificity ranged from 99.7% to 99.8% for all sample types. Overall GC sensitivity ranged from 97.0% to 100% and specificity from 99.9% to 100% see Abstract P3-S1.33 table 1.

Abstract P3-S1.33 Table 1

Sample Type Vag

Sample TypenCT Sensitivity [95% CI]CT Specificity [95% CI]nNG Sensitivity [95% CI]NG Specificity [95% CI]Urine427192.3%(251/272) [88.5 to 94.9]99.7%(3989/3999) [99.5 to 99.9]427498.5%(64/65) [91.8 to 99.7]99.9%(4206/4209) [99.8 to 100]SOV208393.9%(123/131) [88.4 to 96.9]99.7%(1946/1952) [99.3 to 99.9]208397.0%(32/33) [84.7 to 99.5]100%(2049/2050) [99.7 to 100]COV216591.9%(125/136) [86.1 to 95.4]99.8%(2024/20 290) [99.4 to 99.9]2164100%(33/33) [89.6 to 100.0]100%(2130/2131) [99.7 to 100.0]

Conclusions

The cobas assay has excellent sensitivity and specificity when compared to PIS. There was no difference in performance between the SOV and COV specimens or between the vaginal and urine specimens. Self-obtained vaginal swabs provide opportunities for increased efficiency within the clinical settings. The assay is easy to perform, automated, and can be completed in <4 h.
P3-S1.34 Evaluation of the Roche cobas 4800 for the detection ofChlamydia trachomatisandNeisseria gonorrhoeaeusing endocervical specimens
Authors: Pantone A , Van Der Pol B , Lillis R , Loyd B , Nye M B , Taylor S
Published: July 2011

Background

The Roche cobas 4800 (cobas) is a new diagnostic assay that utilises an automated workstation to isolate nucleic acids from clinical specimens and a real time instrument for the detection ofC trachomatis(CT) andN gonorrhoeae(NG). The objective of this study was to compare the performance characteristics of the cobas to the BD Viper (Viper) and GenProbe Aptima Combo 2 (AC2) assays for the detection of CT and NG using a patient infected standard (PIS).

Methods

Specimens were obtained from women attending STD, family planning, or OB/GYN clinics from 12 geographically distinct locations. Endocervical swabs (ES) and samples in ThinPrep liquid based cytology medium (LBC) were obtained from each participant as were urine samples (data not shown) and vaginal swabs (data not shown). LBC were sampled prior to cytology (prequot) for cobas and AC2 and after cytology (postquot) for cobas only. A patient was considered infected if at least 2 of the assays with different molecular targets gave positive results from the ES or urine samples.

Results

Overall CT sensitivity ranged from 89.7 to 92.8% and specificity ranged from 99.6 to 99.8% for all sample types. Overall GC sensitivity ranged from 95.6 to 97.1% and specificity from 99.9 to 100% see Abstract P3-S1.34 table 1.

Abstract P3-S1.34 Table 1

Sample Type Cx

Sample typenCT sensitivity [95% CI]CT specificity [95% CI]nNG sensitivity [95% CI]NG specificity [95% CI]ES425391.6%(240/262) [87.6 to 94.4]99.8%(3984/3991) [99.6 to 99.9]425295.6%(65/68) [87.8 to 98.5]100%(4182/4184) [99.8 to 100]LBC Prequot423892.8%(246/265) [89.1 to 95.4]99.6%(3958/3973) [99.4 to 99.8]423997.1%(67/69) [90.0 to 99.2]99.9%(4167/4170) [99.8 to 100]LBC Postquot420289.7%(235/262) [85.4 to 92.8]99.7%(3930/3940) [99.5 to 99.9]420395.7%(66/69) [88.0 to 98.5]100%(4132/4134) [99.8 to 100.0]

Conclusions

The cobas assay has excellent sensitivity and specificity when compared to PIS. Equivalent performance was observed for the ES and LBC samples, providing clinicians with flexibility to tailor endocervical sample acquisition to their particular setting. There was no statistical difference between the pre- and post-quot LBC samples allowing specimen handling to be suited to the needs of the microbiology and cytology laboratories. The assay is easy to perform, automated, and can be completed in <4 h.
P3-S1.36 Assessing the diagnosis and treatment of urethritis among men attending an urban STD clinic
Authors: Furness B W , Sheriff E A , Sankar S , Elias N Kamanu
Published: July 2011

Background

Urethritis is inflammation of the urethra, the main symptoms of which are dysuria and discharge, and two of the most common causes of which areChlamydia trachomatisandNeisseria gonorrhoeae. The objectives of this study were to: (1) Determine the prevalence of atypical urethritis among participants, (2) Assess the sensitivity and specificity of using gram stain to diagnose gonorrhoea, and (3) Evaluate the effect of diagnostic test used on the time to treatment.

Methods

A random sample of 600 eighteen to 60-year-old men who visited the SE STD Clinic from January 2008 to December 2009 and had a gram stain and nucleic acid amplification test (NAAT) performed were studied. Atypical urethritis was defined as having evidence of inflammation on gram stain but no evidence of gonorrhoea (ie, NGU) and a NAAT negative for both bacteria. The sensitivity and specificity of gram stain were calculated using NAAT as the gold standard. An analysis of variance was used to assess the relationship between time to treatment and diagnostic test—F statistics and corresponding p values were calculated. All statistical analyses were performed using SAS software V. 9 (SAS Institute Inc.) and WinPepi (Abramson, J.H. Epidemiologic Perspective and Innovations).

Results

Of the 600 cases, 493 (82.3%) were Black, 253 (42.2%) had clinical urethritis, and 204 (47.0%) had a previous STD history. The mean age was 30.6 years (SD=10.3). One hundred and five cases of gonorrhoea, 110 cases of chlamydia, and three co-infections were diagnosed (Abstract P3-S1.36 table 1). None of the men diagnosed with gonorrhoea via gram stain were co-infected with chlamydia. The prevalence of atypical urethritis among this study population was 31.2%. The sensitivity of gram stain was 84.3%, the specificity was 100%, the positive predictive value (PPV) was 100%, and the negative predictive value (NPV) was 96.7%. The mean time to treatment was 2.52 days (SD=2.17). Analysis of variance revealed that gram stain (F=41.50, p<0.0001) and NAAT (F=19.18, p<0.0001) had significantly different effects on time to treatment.

Abstract P3-S1.36 Table 1

Concordance between tests used to diagnosis urethritis among men

Gram stain resultNAAT resultTotalGonorrhoeaChlamydiaNegativeGonorrhoea910091NGU3*87*187277*Negative1426195235Total108*113*382600*

Three of these results were positive for both gonorrhoea and chlamydia.

Conclusion

Gram stain is a reliable and relatively inexpensive test for the diagnosis of gonorrhoea among men. Men diagnosed with gonorrhoea or NGU using this point-of-care test were treated earlier than those diagnosed with NAAT only. The prevalence of atypical urethritis among this study population was high and further research should be conducted to investigate the possible aetiologies.
P3-S1.37 The role of microscopy in the diagnosis of proctitis among men who have sex with men (MSM). A retrospective review of rectal screening
Authors: Horner P , Harte D
Published: July 2011

Background

The rectum is a common site of infection withChlamydia trachomatis(CT),Mycoplasma genitalium(MG) andNeisseria gonnorrhoeae(GC) among men who sex with men (MSM) who engage in receptive anal intercourse (RAI). Whilst the majority of individuals remain asymptomatic, these infections are likely to induce rectal inflammation which may have implications for HIV transmission. Information regarding the association between rectal CT, MG and GC and proctitis is limited.

Objectives

To calculate the prevalence of proctitis among a cohort of MSM and investigate the association of proctitis with rectal CT and GC.

Methods

Data were collected retrospectively. All MSM attending the clinic between July and December 2010 were included. Proctitis was defined as >5 PMNLs per high power field on microscopy of a Gram-stained rectal smear. Information including sexual history, clinical presentation, microscopy and GC/CT NAAT (Aptima combo 2 (AC2)) results were recorded on a proforma. Data were analysed using SPSS 16.0. Statistical tests of association were performed using Pearson's χ2test. OR were calculated using logistic regression.

Results

There were 425 MSM who attended for screening in the review period. Of these 83% were UK born. The median age was 32 years (range=16–69 yrs). RAI in the preceding 3 months was reported by 59% (n=251). Of these 64% (n=160) had rectal microscopy performed. Rectal symptoms were reported by 11%. The majority (96%) of samples were collected using a blind rectal swab technique. Proctitis was diagnosed in 23 (14.4%). Of these five were AC2-positive (CT=2, GC=1, CT+GC=1, LGV=1). Rectal AC2 was positive in 19/160 (11.8%) (GC=7, CT=10, CT+GC=1, LGV=1), five of whom had proctitis. Sensitivity and specificity of microscopy for rectal CT/ GC was 26% and 87% respectively. Factors which were predictive of proctitis included rectal symptoms (p=0.001), and being HIV-1sero-positive (p=0.001) but being CT/GC/LGV positive was not associated (p=0.157 OR 2.4; 95% CI=0.8 to 7.5).

Conclusion

Proctitis is common among MSM, particularly HIV-1 seropositive men. The association with GC/CT infection was weak. The causes of and relevance of proctitis remain poorly understood and further research is warranted.
P3-S1.38 Molecular detection ofChlamydia trachomatisandNeisseria gonnorhoeain rectal swabs and (self) collected vaginal swabs with the Cobas 4800 system
Authors: Morre S
Published: July 2011

Objectives

Chlamydia trachomatis(CT) andNeisseria gonnorhoea(NG) are the most prevalent sexual transmitted diseases worldwide. In recent years molecular diagnostics for the standard sample types like cervical swabs and urines has been highly adapted in most laboratories. Currently more difficult samples types, with regard to inhibition, sensitivity and specificity, like rectal swabs and (self collected) vaginal swabs are received more often in the laboratories. This study describes the performance of the new Cobas 4800 for the detection of CT and NG in these difficult sample types.

Methods

A total of 1100 co-collected swabs were tested (900 vaginal; 200 rectal). Informed consent was obtained from all patients included in the study for co-collection. All swabs were tested routinely with the M2000 system (Abbott) and the co-collected samples were tested in the Cobas 4800 system (Roche). All swabs were collected in their corresponding transport buffer and tested blinded. Discrepant results were tested with an independent real time PCR method.

Results

In the 900 vaginal swabs 10% (n=90) were positive for CT and 0.5% (n=4) for NG and in the 200 rectal swabs 11% CT positives (n=22) an 6% NG positives (n=12) were identified. In the vaginal samples nine discrepant results between the Cobas 4800 and M2000 system were identified. Five with borderline values and four with clear different results (1 NG, 3CT). Two M2000 positive results (Ct 35.9; 32.3) were available for home brew analysis and could not be confirmed. In the rectal samples 8 discrepant results were found including three clear differences (2 NG; 1 CT). The CT discrepant sample was positive in the Cobas 4800 system (Cp 33.1) and negative in the M2000 system. For NG two samples were negative in the Cobas 4800 system but positive (Ct 23.8; 32.0) in the M2000 system. All three clear discrepant results were available for home brew analysis and confirmed the Cobas 4800 results.

Conclusion

Both samples types showed a high concordance between the two systems (ĸ 0.95 for CT; ĸ 0.93 for NG). For detection of vaginal swabs some small differences were found both for CT and NG but these were equally spread between the two systems. There is no difference between self collected and clinician taken vaginal swabs. All clear rectal swabs discrepancies confirmed the Cobas 4800 results. This remarkable difference has to be further studied. In general, (self collected) vaginal swabs and rectal swabs show reliable results for routine detection of CT and NG.
P1-S1.44 Antimicrobial susceptibility profile ofNeisseria gonorrhoeaeisolates in the Province of QuÉbec - 2010
Authors: Lefebvre B , Bourgault A M
Published: July 2011

Background

Regular monitoring of antimicrobial resistance ofN gonorrhoeaeis necessary to detect the trend in antimicrobial resistance, increases in minimum inhibitory concentrations (MICs) and to assist in the development of treatment guidelines.

Methods

In 2010, allN gonorrhoeaestrains isolated in clinical laboratories throughout the province were submitted to the Laboratoire de santé publique du Québec where their susceptibility profile to azithromycin, cefixime, ceftriaxone, ciprofloxacin and spectinomycin was determined by the agar dilution method.

Results

A total of 831 strains isolated from 607 males and 224 females were tested. The strains were recovered from (data available for 819 strains) the following specimens—urethra, 432; cervix, 191; anus, 100; throat, 90; eye, 4; and synovial fluid, 2. All strains were susceptible to cefixime, ceftriaxone and spectinomycin, 270 (32.5%) were resistant to ciprofloxacin and 11 (1.3%) were resistant to azithromycin (MIC ≥2 mg/l). The azithromycin resistant isolates were retrieved from 11 males aged 16–55 years (mean, 30) from the Montreal area (urethra, 8; throat, 3). The azithromycin MIC distribution was as follows—4 mg/l (n=1), 8 mg/l (n=6), 16 mg/l (n=4). No highly resistant organism (MIC>128 mg/l) was identified. Among the azithromycin resistant isolates, one was resistant to ciprofloxacin but all were susceptible to cefixime, ceftriaxone and spectinomycin. In 2010, 68 (8.2%) isolates exhibited decreased susceptibility to cefixime (MIC=0.125 mg/l [n=62] and 0.25 mg/l [n=6]). Of these 68 isolates, all were susceptible to azithromycin and ceftriaxone but all were resistant to ciprofloxacin. Only one isolate showed decreased susceptibility to ceftriaxone (MIC=0.125 mg/l)—this isolate also showed decreased susceptibility to cefixime.

Conclusions

As observed elsewhere,N gonorrhoeaestrains are showing a worrisome drift in susceptibility to cefixime, a first line treatment for gonococcal infections associated with resistance to ciprofloxacin. Furthermore, resistance to azithromycin, a second line treatment option, is emerging. Sustained collaborative laboratory surveillance programs are mandatory for the design of public health interventions to prevent and control gonococcal infections.
P3-S1.41 Coinfection ofNeisseria gonorrhoeaeandChlamydia trachomatisin symptomatic and asymptomatic women in India: implications in reproductive health
Authors: Sachdev D , Sonkar S , Mishra P , Patel A , Saluja D
Published: July 2011

Background

Despite the recent advances in diagnosis,Neisseraia gonorrhoeae(NG) andChlamydia trachomatis(CT) remain leading cause of STDs worldwide and account for STI's morbidity leading to acquisition of HIV and HPV infection. Coinfection of gonococcus and Chlamydia is reported from various countries. In recent study we reported high rate of infection by CT using an in house developed PCR method. A number of studies also suggest increasing rate of antibiotic resistance in NG. Thus we developed a rapid, specific and cost effective diagnostic method to detect prevalence of co infection by NG and CT. Methodology: (1) Unique gene sequence of CT and NG were amplified from gDNA isolated from endocervical swabs. (2) Validation of in house PCR method using Roche AMPLICOR Micro well plate CT/NG kit. (3) Use of molecular beacon to develop easy visual method. (4) Establishment of multiplex PCR (mPCR) for simultaneous detection of CT and NG.

Results

360 clinical samples were used to validate in house PCR assay. Discrepancy of the samples was resolved by amplifying genes encoding for outer membrane proteins (rmp for NG and ompA for CT). The resolved PPV and NPV were found to be 94% and 99% for CT, 92.0% and 96% for NG. Molecular beacon probe was used which helped in visualisation of PCR product directly in tube using dark reader which also improved the sensitivity of assay. The overall infection rate by NG was 26% and 8.6% in symptomatic and asymptomatic patients while that of CT was 26.3% and 21% respectively. To make the method easy to use in remote areas with minimum laboratory infrastructure, the in house PCR assay was standardised for detection using dry swabs, with crude DNA preparation and stabilisation of reagents at 4C (up to 6 months) was achieved for easy transportation and storage. Using single PCRs, coinfection by CT and NG was found to be 18% in symptomatic (74/410) and 5 % in asymptomatic patients (18/360). We further developed mPCR for simultaneous detection two pathogens. Sensitivity of in house mPCR was found to be 95% when evaluated against PCR for individual pathogen.

Conclusion

Detection of both the pathogens in single PCR assay makes it economical both in terms of cost and time. The in house assay is highly sensitive, easy to perform and requires minimum infrastructure as well as technical expertise, making it a better option for routine diagnosis of genital infection in developing countries, which would be of great consequences in disease management.
P3-S1.42 Complex treatment of urogenital chlamydiosis with macrolids and immunostimulants
Authors: Fartushok T , Dasyuk T
Published: July 2011

Objectives

The data concerning treatment of such sexually transmitted infection asChlamydia trachomatis(CTR) among Ukrainian population are still limited and controversial. This information is of great scientific and health care interest. The aim of investigation is to find out efficiency of urogenital chlamydiosis treatment involving new effective ways of treatment and choose the most appropriate and expedient method of treatment, which would completely meet recommendations of WHO.

Methods

We have analysed literature, records and recommendations concerning treatment of patients with chlamydial infection and concluded that there is no a perfect method of treatment, which would ensure complete clinical and laboratory cure. According to literature data, macrolids are good drugs with insignificant resistance to agent, nevertheless, recovery of a patient is, sometimes, incomplete due to complications. We have examined patients with urogenital chlamydiosis. All patients (n=134) were divided into four groups and treated with antimicrobial drugs jozamicyn and azytromicyn, and immunomodulators - manax and kagotsel. Patients from the 1st group (n=35) were treated with jozamicyn and manax, 2nd group (n=35) with azytromicyn and manax, 3rd group (n=34) with jozamicyn and kagotsel, 4th group (n=30) with azytromicyn and kagotsel.

Results

Following treatment results of our investigation showed complete laboratory and clinical recovery in 93.2 % of patients from the 1st group, 98.1% of patients from the 2nd group, 91.8% of patients from the 3rd group, and 93.5% of patients from the 4th group.

Conclusion

According to obtained data, our method of treatment, which was used for treatment of patients from the 2nd group, is the most appropriate for common application in the treatment of urogenital chlamydiosis