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ABSTRACT: A polymerase chain reaction (PCR)-based enzyme-linked immunosorbent assay (ELISA) methodology was developed to characterize Neisseria gonorrhoeae porB gene variable regions (VR); the methodology was evaluated in comparison to porB VR typing by checkerboard hybridization. Clinical noncultured samples from 35 men who have sex with men (MSM), positive by nucleic amplification assays for N. gonorrhoeae, were typed using a panel of 40 oligonucleotide probes to porB VRs and compared to checkerboard hybridization. Complete concordance was observed between the two methods at PIB VRs 1, 3, and 7. At the more degenerate VRs 5 and 6, PCR ELISA resulted in obtaining more typeable VRs than checkerboard hybridization due to single nucleotide mismatches. By PCR ELISA, two predominant PIB porB types were identified in 58% of the samples and the remaining 16 samples had one of six other porB types. Both PCR ELISA and checkerboard hybridization methods of porB VR typing allowed characterization of N. gonorrhoeae from noncultured clinical samples including throat and rectal swabs and discriminated N. gonorrhoeae from N. meningitidis present in some of the samples. PCR ELISA is a rapid, relatively inexpensive and alternative molecular typing method for N. gonorrhoeae, suitable for use in conjunction with molecular diagnostic tests.
Journal of Clinical Laboratory Analysis 02/2007; 21(4):237-43. · 1.38 Impact Factor
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ABSTRACT: Our aim was to determine the incidence of re-infection of men who have sex with men (MSM) for rectal Neisseria gonorrhoeae (RNg) and rectal Chlamydia trachomatis (RCt) infection. The medical records of 126 MSM diagnosed with RNg or RCt infections at Melbourne Sexual Health Centre were reviewed. A total of 68 of the 126 (54%) MSM were re-tested. Among the HIV-positive MSM, 17 (81%) of 21 were re-tested and eight were positive (47%, 95% confidence interval [CI] 23-72%). Among the 51 (49%) of 105 of the HIV-negative or unknown MSM re-tested, 13 (25%, 14-40) were positive. Of the 21 infections identified, five cases were symptomatic. The incidence of rectal infections among MSM is high and prior RNg or RCt should be included as a risk factor for frequent screening.
International Journal of STD & AIDS 07/2006; 17(6):415-7. · 1.09 Impact Factor
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ABSTRACT: Guidelines for testing men who have sex with men (MSM) were published in 2002. They did not recommend asymptomatic screening for urethral gonorrhoea or pharyngeal screening for chlamydia. To determine if these guidelines were appropriate, we audited gonorrhoea and chlamydia testing of MSM at our Centre.
We carried out two audits at our Centre between August 2001 and July 2002. The first was an audit of testing MSM for gonorrhoea and/or chlamydia over 12 days. The second was an audit of all positive tests over this 12-month period for gonorrhoea or chlamydia among MSM.
During the 12 selected days 89 of 286 men tested (31%) were MSM. Among the MSM testing positive for gonorrhoea and/or chlamydia infection (15, 17%), symptomatic urethral infection was the most common (n = 8). No rectal and pharyngeal infections had site-specific symptoms. Based on the guidelines, 100 of the 334 tests ordered (30%) were not recommended according to the guidelines, and none of these 100 tests yielded a positive result. Over the 12-month audit period, 135 MSM were diagnosed with gonorrhoea and/or chlamydia. For gonorrhoea, site specific symptoms were present in 42 of 43 cases of urethral infection (98%), six of 23 cases of rectal infection (26%), and no cases of pharyngeal infection had symptoms. For chlamydia, site-specific symptoms were present in 29 of 48 cases of urethral infection (60%), six of 33 cases of rectal infection (18%), and in one of the two cases of pharyngeal chlamydia identified. A substantial proportion of cases occurred in clients with HIV infection (21, 16%).
These findings strongly support screening among MSM and in particular not testing asymptomatic MSM for urethral gonorrhoea or any MSM for pharyngeal chlamydia.
Sexual Health 02/2004; 1(1):47-50. · 1.45 Impact Factor
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ABSTRACT: Screening guidelines for men who have sex with men (MSM) recommend testing of extragenital sites (pharyngeal and rectal) for gonorrhoea and chlamydia. Testing of specimens from these sites is not validated by most commercial nucleic amplification tests, such as the COBAS Amplicor assay. To investigate the utility of the COBAS Amplicor assay for detection of Chlamydia trachomatis in extragenital specimens, this study developed and evaluated confirmatory tests using the omp1 gene as an alternative target for amplification by PCR. Of anal and throat swabs collected from men in male-only saunas, 52 swabs that tested C. trachomatis positive by COBAS Amplicor and 30 swabs that tested as negative were included for confirmatory omp1 PCR testing. A total of 49 (94%) COBAS Amplicor-positive samples were confirmed by the omp1 PCR. A substantial proportion of specimens were confirmed by using a nested omp1 PCR (27%). Not confirmed by any omp1 PCR were three anal swabs (6%). It is most probable that these samples contained lower bacterial levels that were near or below the detection level of the omp1 PCR assays. The findings of this study support the confident reporting of C. trachomatis detected by COBAS Amplicor in extragenital specimens and support the utility of this assay as a screening test for MSM.
Journal of Clinical Microbiology 02/2004; 42(1):239-41. · 4.15 Impact Factor
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ABSTRACT: In response to increases in sexually transmissible infections (STI) and HIV infection rates among men who have sex with men (MSM), the current study aimed to investigate the feasibility of a screening program at male-only saunas in Melbourne, Australia.
The goal was to determine (1) the participation rate, and the proportion whom obtain test results; (2) the prevalence of gonorrhea and chlamydia; and (3) to evaluate risk factors for STI acquisition.
We used a cross-sectional design. Pharyngeal, rectal, and urethral specimens were collected from participants, and tested for Chlamydia trachomatis and Neisseria gonorrhoeae by polymerase chain reaction (PCR).
There was a participation rate of 24% (n=521), and 70% obtained their test results. The infection rate in those who failed to collect their results was no different than those seeking theirs. The proportion of participants with PCR-detected gonorrhea and/or chlamydia infection was high, 10.7%. The presence of infection was associated with seeking sexual health care in the last year.
The high prevalence rate of gonorrhea and chlamydia supports the concept of a screening program in Melbourne male-only saunas. The low participation rate has highlighted the need to consider alternative methods for making contact with men in the saunas or offering incentives to participate in future screening programs. Although anonymous participation encouraged participation for some men, future programs should attempt to obtain contact details for follow up of positive test results.
Sex Transm Dis 01/2004; 30(12):886-9. · 2.87 Impact Factor
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Sexually Transmitted Infections 03/2003; 79(1):78. · 2.85 Impact Factor
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Sexually Transmitted Infections 11/2002; 78(5):387-8. · 2.85 Impact Factor
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J Clin Microbiol. 43(5):2546-7.
