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Molecular methods that characterize the Neisseria gonorrhoeae porin protein Por are needed to study gonococcal pathogenesis in the natural host and to classify strains from direct clinical samples used with nucleic acid amplification-based tests. We have defined the capabilities of por variable region (VR) typing and determined suitable conditions to apply the method to direct clinical specimens. Nested PCR from spiked urine samples detected 1 to 10 copies of template DNA; freezing spiked whole urine greatly reduced the ability to amplify porB . In a laboratory model of mixed gonococcal infections, the por type of one strain could be determined in the presence of a 100-fold excess of another. por VR typing was used to examine clinical samples from women enrolled in studies conducted in Baltimore, Md., and Madagascar. por type was determined from 100% of paired cervical swab and wick samples from 20 culture-positive women from Baltimore; results for eight individuals (40%) suggested infection with more than one strain. In frozen urine samples from Madagascar, porB was amplified and typed from 60 of 126 samples from ligase chain reaction (LCR)-positive women and 3 samples from LCR-negative women. The por VR types of 13 samples (21%) suggested the presence of more than one gonococcal strain. Five por types, identified in >45% of women with typed samples, were common to both geographic areas. Molecular typing is an important adjunct to nucleic acid amplification-based diagnostics. Methods that utilize direct clinical samples and can identify mixed infections may contribute significantly to studies of host immunity, gonococcal epidemiology, and pathogenesis.

Objectives: Trichomonas vaginalis is the most common nonviral sexually transmitted pathogen. Wet mount microscopy performs poorly compared with culture; however, universal screening using culture would be cost-prohibitive. Goal: The goal of this study was to develop a predictive model for wet mount-negative women who may benefit from targeted use of culture for T. vaginalis detection. Study: Women presenting for sexually transmitted disease evaluation were prospectively screened for trichomoniasis using wet mount and culture. Multivariate logistic regression was used to identify predictors of culture-proven trichomoniasis among wet mount-negative women. Results: A total of 2194 women were screened for trichomoniasis; overall, the prevalence of T. vaginalis was 17.5%. Three predictors (any drug use, contact to trichomoniasis, and African-American race) provided the most specific model (100%); any 1 predictor, the most sensitive model (97.8%). Conclusions: Given the public health impact of trichomoniasis, we recommend using any 1 of 3 predictors to identify women who would benefit from targeted culture for T. vaginalis.

Variations in estimates of prevalence of trichomoniasis in men may reflect true differences in the burden of disease but are also affected by the performance of diagnostic methods and the type of specimen tested. In this study, men were evaluated at baseline and at follow-up, to evaluate syndromic management of urethritis and the effects of human immunodeficiency virus and trichomoniasis, in Lilongwe, Malawi. First-void urine specimens and urethral swabs were obtained at enrollment, for Trichomonas vaginalis culture; semen specimens were also obtained at follow-up. The sensitivities of testing methods using urine specimens and urethral swabs were equal; 67% of cases were identified by use of either specimen, and, in 47% of cases, both specimens tested positive. When semen specimens were included, all 3 specimens tested positive in only 19% of cases. Semen was the most sensitive single specimen, and, in 25.6% of cases, only semen specimens tested positive. Thus, prevalence of T. vaginalis infection in men is underestimated if only 1 specimen is tested.

Trichomonas vaginalis was originally considered a commensal organism until the 1950s when the understanding of its role as a sexually transmitted infection (STI) began to evolve. Trichomoniasis has been associated with vaginitis, cervicitis, urethritis, pelvic inflammatory disease (PID), and adverse birth outcomes. Infection with T vaginalis could have an important role in transmission and acquisition of HIV. T vaginalis is site specific for the genitourinary tract and has been isolated from virtually all genitourinary structures. Asymptomatic disease is common in both men and women, thus screening for disease is important. Various sociodemographic factors have been correlated with presence of T vaginalis, and may be used to predict infection. Diagnosis is usually made from wet mount microscopy and direct visualisation, which are insensitive. DNA amplification techniques perform with good sensitivity, but are not yet approved for diagnostic purposes. In areas where diagnostic methods are limited, management of trichomoniasis is usually as part of a clinical syndrome; vaginal discharge for women and urethral discharge for men. A single dose of metronidazole is effective in the majority of cases. Outside of the United States, other nitroimidazoles may be used and are as effective as metronidazole. Metronidazole resistance is an emerging problem, but its clinical importance is not yet clear. Concomitant treatment of sexual partners is recommended.

Background: Male urethritis is generally treated syndromically but failure of empirical treatment is common. Goal: The study goal was to evaluate the addition of metronidazole to the syndromic management of urethritis in Malawi in a randomized clinical trial. Study Design: Men with urethritis were randomized to receive either 2 g of metronidazole by mouth or placebo in addition to standard care for urethritis (i.e. a single intramuscular dose of 240 mg gentamicin and 100 mg doxycycline twice daily for 7 days). The primary endpoints of the study included measurement of the effects of treatment on Trichomonas vaginalis signs and symptoms of urethritis and the concentration of HIV RNA in semen in dually infected subjects. Results: The overall prevalence of T vaginalis was 17.3% (71/411) and treatment with metronidazole cleared 95% of culture-positive infections compared with 54% clearance among men receiving placebo (P = 0.006). Prevalence of persistent urethritis was observed in approximately 16% of both groups at the end of 1 week (29/179 of those receiving metronidazole versus 29/187 in the placebo group; P = 0.86). For a subset of HIV-infected men with trichomoniasis the seminal plasma HIV RNA concentration was lower than in a group of HIV-positive control subjects (P = 0.052). Conclusion: In areas with a high prevalence of trichomoniasis the addition of metronidazole to the syndromic management of male urethritis can eliminate infection with T vaginalis and may help to reduce the transmission of HIV. Such treatment should be strongly considered as part of empirical therapy for urethritis in men in Malawi and places where T vaginalis infection in men is common. (authors)

Modification of the standard Chlamydia trachomatis ligase chain reaction (LCR) detection assay resulted in a quantitative test. Sample rates from C. trachomatis standards ranging from 32 to 1,048,576 elementary bodies (EB)/ml of urine exhibited a linear correlation with concentration. Quantitative LCR (qLCR) was used to measure the number of EB per milliliter in 158 urine samples from women in Madagascar that tested positive for C. trachomatis by the standard LCR detection assay. C. trachomatis concentrations displayed an apparent bimodal distribution, with approximately one-third of samples (37%) in a peak ranging from 32 to 1,015 EB/ml (median = 297 EB/ml) and the remainder (63%) in a grouping with relatively higher concentrations, ranging from 1,086 to 218,670 EB/ml (median = 7,389 EB/ml). qLCR will be useful for studies of chlamydial infection aimed at understanding the associations of organism burden with clinical manifestations and transmission.

Trichomonas vaginalis (TV) is a common sexually transmitted infection that can cause vaginitis, cervicitis and urethritis. Persistent and recurrent TV infections are frequent in women, potentially due to the lack of routine screening recommendations for this pathogen, the chronic nature of some infections, and drug resistance. Metronidazole and tinidazole are two oral drugs that are effective against trichomoniasis. There are few alternative treatment options for persons with a metronidazole allergy or treatment failure. Most TV isolates from women with treatment failures that have been analyzed for susceptibility testing in the United States have exhibited low-level metronidazole resistance, supporting the initial use of tinidazole for patients who fail metronidazole therapy. Several non-nitroimidazole drugs and other agents have demonstrated acceptable in vitro activity or cure rates in case reports for metronidazole-resistant trichomoniasis; however, clinical trials are imperative to evaluate their efficacy as alternative therapeutic regimens for this highly prevalent infection.

A biomarker of unprotected receptive anal intercourse could improve validity of sexual behavior measurement. We quantified prostate-specific antigen (PSA) from rectal swabs from men who have sex with men (MSM). One swab was PSA positive. Using current methods, PSA is an inadequate biomarker of recent unprotected receptive anal intercourse in men who have sex with men.

T. vaginalis infection (trichomoniasis) is the most common curable sexually transmitted infection (STI) in the U.S. It is associated with increased HIV risk and adverse pregnancy outcomes. Trichomoniasis surveillance data do not exist for either national or local populations. The Monitoring STIs Survey Program (MSSP) collected survey data and specimens which were tested using nucleic acid amplification tests to monitor trichomoniasis and other STIs in 2006–09 among a probability sample of young adults (N = 2,936) in Baltimore, Maryland — an urban area with high rates of reported STIs. The estimated prevalence of trichomoniasis was 7.5% (95% CI 6.3, 9.1) in the overall population and 16.1% (95% CI 13.0, 19.8) among Black women. The overwhelming majority of infected men (98.5%) and women (73.3%) were asymptomatic. Infections were more common in both women (OR = 3.6, 95% CI 1.6, 8.2) and men (OR = 9.0, 95% CI 1.8, 44.3) with concurrent chlamydial infection. Trichomoniasis did not vary significantly by age for either men or women. Women with two or more partners in the past year and women with a history of personal or partner incarceration were more likely to have an infection. Overall, these results suggest that routine T vaginalis screening in populations at elevated risk of infection should be considered.

Background: Ebola virus (EBOV) RNA persistence in semen, reported sexual transmission, and sporadic clusters at the end of the 2013-2016 epidemic have prompted recommendations that male survivors refrain from unprotected sex unless their semen is confirmed to be EBOV free. However, there is no fully validated assay for EBOV detection in fluids other than blood. Methods: The Cepheid Xpert Ebola assay for EBOV RNA detection was validated for whole semen and blood using samples obtained from uninfected donors and spiked with inactivated EBOV. The validation procedure incorporated standards from Clinical and Laboratory Standards Institute and Good Clinical Laboratory Practices guidelines for evaluating molecular devices for use in infectious disease testing. Results: The assay produced limits of detection of 1000 copies/mL in semen and 275 copies/mL in blood. Limits of detection for both semen and blood increased with longer intervals between collection and testing, with acceptable results obtained up to 72 hours after specimen collection. Conclusions: The Cepheid Xpert Ebola assay is accurate and precise for detecting EBOV in whole semen. A validated assay for EBOV RNA detection in semen informs the care of male survivors of Ebola, as well as recommendations for public health.

Trichomonas vaginalis is one of the most common sexually transmitted infections. In Malawi, rates of trichomoniasis in women are high. The prevalence of T. vaginalis infection in men is expected to be high but has not previously been documented.We sought to determine the prevalence of trichomoniasis in Malawian men with and without urethritis, to evaluate a polymerase chain reaction detection assay for T. vaginalis in urethral swabs and to examine the effect of T. vaginalis infection on excretion of human immunodeficiency virus (HIV) in semen.Men presenting at the Sexually Transmitted Diseases (STD) and Dermatology Clinics in Malawi were enrolled in a cross-sectional study. We compared a polymerase chain reaction-based test for T. vaginalis detection with wet-mount microscopy and culture of urethral swabs. HIV serology was determined by enzyme-linked immunosorbent assay (ELISA), and HIV-1 RNA concentrations in semen were measured by quantitative nucleic acid sequence-based analysis.T. vaginalis was detected in 51 of 293 men. The estimated prevalence among symptomatic men was 20.8% and among asymptomatic men, 12.2%. Polymerase chain reaction performed with a sensitivity of 0.82 (95% CI: 0.66-0.92) and specificity of 0.95 (95% CI: 0.91-0.97) compared to wet-mount microscopy and culture. There was no difference in the rate of HIV seropositivity in men with and without T. vaginalis infection. However, in men with symptomatic urethritis, the median HIV RNA concentration in seminal plasma from men with T. vaginalis was significantly higher that in seminal plasma from HIV-positive men without trichomonas.This study was conducted to determine the prevalence of trichomoniasis in Malawian men, to evaluate a polymerase chain reaction (PCR) detection assay for T. vaginalis in urethral swab samples, and to examine the effect of T. vaginalis infection on HIV excretion in the semen. There were 206 men with symptomatic urethritis in STD clinic and 127 asymptomatic men in the Dermatology Clinic who were enrolled from January to March 1996. Results, according to a wet-mount microscopy and urethral swabs culture combination, showed that, of 293 men, only 38 (13%) men were positive for T. vaginalis. The estimated prevalence among symptomatic and asymptomatic cases was 15.7% and 8.7%, respectively. The PCR yielded a sensitivity of 0.82 (95% CI: 0.66-0.92) and specificity of 0.95 (95% CI: 0.91-0.97); these were compared to the wet-mount microscopy and culture combination. Overall HIV seroprevalence of men was 51%, because gonococcal urethritis was shown to significantly increase seminal HIV RNA levels. The median HIV RNA concentration in seminal plasma from men with symptomatic urethritis plus T. vaginalis infection was significantly higher than in seminal plasma from HIV-positive men with symptomatic urethritis only. Since this study has several important limitations, a randomized clinical trial would be useful for determining whether urethritis cure rates can be significantly improved.

Phosphoethanolamine (PEA) on Neisseria gonorrhoeae lipid A influences gonococcal inflammatory signaling and susceptibility to innate host defenses in in vitro models. Here, we evaluated the role of PEA-decorated gonococcal lipid A in competitive infections in female mice and in male volunteers. We inoculated mice and men with mixtures of wild-type N. gonorrhoeae and an isogenic mutant that lacks the PEA transferase, LptA. LptA production conferred a marked survival advantage for wild-type gonococci in the murine female genital tract and in the human male urethra. Our studies translate results from test tube to animal model and into the human host and demonstrate the utility of the mouse model for studies of virulence factors of the human-specific pathogen N. gonorrhoeae that interact with non-host-restricted elements of innate immunity. These results validate the use of gonococcal LptA as a potential target for development of novel immunoprophylactic strategies or antimicrobial treatments. IMPORTANCE Gonorrhea is one of the most common bacterial sexually transmitted infections, and increasing antibiotic resistance threatens the use of currently available antimicrobial therapies. In this work, encompassing in vitro studies and in vivo studies of animal and human models of experimental genital tract infection, we document the importance of lipid A’s structure, mediated by a single bacterial enzyme, LptA, in enhancing the fitness of Neisseria gonorrhoeae. The results of these studies suggest that novel agents targeting LptA may offer urgently needed prevention or treatment strategies for gonorrhea., IMPORTANCE Gonorrhea is one of the most common bacterial sexually transmitted infections, and increasing antibiotic resistance threatens the use of currently available antimicrobial therapies. In this work, encompassing in vitro studies and in vivo studies of animal and human models of experimental genital tract infection, we document the importance of lipid A’s structure, mediated by a single bacterial enzyme, LptA, in enhancing the fitness of Neisseria gonorrhoeae. The results of these studies suggest that novel agents targeting LptA may offer urgently needed prevention or treatment strategies for gonorrhea.

Research on interventions to prevent HIV and other sexually transmitted infections (STIs) is heavily influenced by participant reporting of sexual behavior, despite uncertainty about its validity. Exclusive reliance on participant self-report often is based, overtly or by implication, on 4 assumptions: (1) no feasible alternatives exist; (2) misreporting can be minimized to levels that can be disregarded; (3) misreporting tends to underreport sensitive behaviors; and (4) misreporting tends to be nondifferential with respect to the groups being compared. The objective of this review are to evaluate these assumptions, including a review of studies using semen biomarkers to evaluate the validity of self-reported data, and to make recommendations for applying biological markers of semen exposure detectable in women to further strengthen research on HIV/STI prevention. Increasing evidence shows that semen biomarkers provide an important means of assessing and augmenting the validity of studies on HIV/STI prevention. Additional biomarkers are needed to assess male exposure to vaginal sex and both male and female exposure to anal sex. Methods and study designs that incorporate biomarkers into studies collecting self-reported behavioral data should be considered where possible.

Recent advances in tests for the sexually transmitted protozoan parasite Trichomonas vaginalis have increased opportunities for diagnosis and treatment of this important sexually transmitted infection. This review summarises currently available tests, highlighting their performance characteristics, advantages and limitations. The recent development of molecular tests for the detection of T vaginalis, including rapid antigen detection and nucleic acid amplification tests, has significantly improved the quality of diagnostics for trichomoniasis, particularly in women. In light of the expanded menu of testing options now available, improved recognition and better control of trichomoniasis are in sight, which should enable the eventual reduction of adverse reproductive consequences associated with T vaginalis infection.

BackgroundHuman immunodeficiency virus (HIV) prevalence among men who have sex with men (MSM) is thought to be high in Jamaica. The objective of this study was to estimate HIV prevalence and identify risk factors in order to improve prevention approaches.MethodsWith the help of influential MSM, an experienced research nurse approached MSM in four parishes to participate in a cross-sectional survey in 2007. Men who have sex with men were interviewed and blood taken for HIV and syphilis tests, and urine taken for gonorrhoea, chlamydia and trichomonas testing using transcription-mediated amplification assays. A structured questionnaire was administered by the nurse.ResultsOne third (65 of 201; 32%, 95% Confidence Interval (CI) 25.2, 47.9) of MSM were HIV positive. Prevalence of other sexually transmitted infections (STI) was: chlamydia 11%, syphilis 6%, gonorrhoea 3.5% and trichomonas 0%. One third (34%) of MSM identified themselves as being homosexual, 64% as bisexual and 1.5% as heterosexual. HIV positive MSM were significantly more likely to have ever been told by a doctor that they had an STI (48% vs 27%, OR 2.48 CI 1.21, 5.04, p = 0.01) and to be the receptive sexual partner at last sex (41% vs 23%, OR 2.41 CI 1.21, 4.71, p = 0.008). Men who have sex with men who were of low socio-economic status, ever homeless and victims of physical violence were twice as likely to be HIV positive. The majority (60%) of HIV positive MSM had not disclosed their status to their partner and over 50% were not comfortable disclosing their status to anyone.ConclusionsThe high HIV prevalence among MSM is an important factor driving the HIV epidemic in Jamaica. More effective ways need to be found to reduce the high prevalence of HIV among MSM including measures to reduce their social vulnerability, combat stigma and discrimination and empower them to practice safe sex.

A physical and rudimentary genetic map of the Haemophilus ducreyi strain 35000 genome was constructed. Pulsed-field gel electrophoresis was used to separate restriction fragments of H. ducreyi DNA digested with Sfil, I-Ceul, or Sfil plus I-Ceul. The sizes of the fragments were determined, and the circular chromosome was estimated to be 1757 kbp. The six I-Ceul fragments and four Sfil fragments were ordered into macrorestriction maps using Southern blot hybridization with random H. ducreyi clones as probes. It was shown that both H. ducreyi and the distantly related Haemophilus influenzae have six rrn operons marked by the locations of the I-Ceul sites. However, the two species displayed distinct I-Ceul restriction patterns. A second H. ducreyi strain, CIP542, displayed an identical I-Ceul pattern to that of H. ducreyi 35000, but Sfil digests of teh two strains were distinct. The orientation of the six rrn operons was determined and thirteen identified H. ducreyi genes positioned on the map of strain 35000.

Haemophilus ducreyi is a strict human pathogen that causes sexually transmitted genital ulcer disease. We infected domestic swine with H. ducreyi 35000, resulting in the development of cutaneous ulcers histologically resembling human chancroid lesions. Intraepidermal lesions progressed from pustules to ulcers containing polymorphonuclear leukocytes and were accompanied by a dermal inflammatory infiltrate containing T cells and macrophages. H. ducreyi was recovered from lesions up to 17 days after inoculation, and pigs did not develop immunity to reinfection with the challenge strain. Features of the model include inoculation through abrasions in the epidermis, ambient housing temperatures for infected pigs, the ability to deliver multiple different inocula to a single host, and the availability of monoclonal antibodies against porcine immune cells permitting immunohistochemical characterization of the host immune response to H. ducreyi infection.

Trichomoniasis is a common sexually transmitted disease associated with preterm birth, low birth weight, and increased susceptibility to infection with other pathogenic sexually transmitted microorganisms. Nucleic acid amplification tests for Trichomonas vaginalis have improved sensitivity for detecting infected individuals compared to existing culture-based methods. This prospective, multicenter U.S. clinical trial evaluated the performance of the automated Aptima T. vaginalis assay for detecting T. vaginalis in 1,025 asymptomatic and symptomatic women. Vaginal swab, endocervical swab, ThinPrep PreservCyt, and urine specimens were collected. Subject infection status was determined by wet-mount microscopy and culture. Aptima T. vaginalis assay performance was determined for each specimen type by comparison to subject infection status. Of 933 subjects analyzed, 59.9% were symptomatic. Aptima T. vaginalis clinical sensitivity and specificity were, respectively, 100% and 99.0% for vaginal swabs, 100% and 99.4% for endocervical swabs, 100% and 99.6% in ThinPrep samples, and 95.2% and 98.9% in urine specimens. Aptima T. vaginalis performance levels were similar in asymptomatic and symptomatic subjects. This study validates the clinical performance of the Aptima T. vaginalis assay for screening asymptomatic and symptomatic women for T. vaginalis infection.

Haemophilus ducreyi , the etiologic agent of chancroid, has an obligate requirement for heme. Heme is acquired by H. ducreyi from its human host via TonB-dependent transporters expressed at its bacterial surface. Of 3 TonB-dependent transporters encoded in the genome of H. ducreyi , only the hemoglobin receptor, HgbA, is required to establish infection during the early stages of the experimental human model of chancroid. Active immunization with a native preparation of HgbA (nHgbA) confers complete protection in the experimental swine model of chancroid, using either Freund's or monophosphoryl lipid A as adjuvants. To determine if transfer of anti-nHgbA serum is sufficient to confer protection, a passive immunization experiment using pooled nHgbA antiserum was conducted in the experimental swine model of chancroid. Pigs receiving this pooled nHgbA antiserum were protected from a homologous, but not a heterologous, challenge. Passively transferred polyclonal antibodies elicited to nHgbA bound the surface of H. ducreyi and partially blocked hemoglobin binding by nHgbA, but were not bactericidal. Taken together, these data suggest that the humoral immune response to the HgbA vaccine is protective against an H. ducreyi infection, possibly by preventing acquisition of the essential nutrient heme.

Experimental infection of male volunteers with Neisseria gonorrhoeae is safe and reproduces the clinical features of naturally acquired gonococcal urethritis. Human inoculation studies have helped define the natural history of experimental infection with two well-characterized strains of N. gonorrhoeae, FA1090 and MS11mkC. The human model has proved useful for testing the importance of putative gonococcal virulence factors for urethral infection in men. Studies with isogenic mutants have improved our understanding of the requirements for gonococcal LOS structures, pili, opacity proteins, IgA1 protease and the ability of infecting organisms to obtain iron from human transferrin and lactoferrin during uncomplicated urethritis. The model also presents opportunities to examine innate host immune responses that may be exploited or improved in development and testing of gonococcal vaccines. Here we review results to date with human experimental gonorrhea.

Trichomonas vaginalis (TV) infection is the most prevalent curable sexually transmitted infection in the United States and worldwide. Most TV infections are asymptomatic, and the accurate diagnosis of this infection has been limited by lack of sufficiently sensitive and specific diagnostic tests, particularly for men. To provide updates for the 2010 Centers for Disease Control and Prevention’s Sexually Transmitted Diseases Treatment Guidelines, a PubMed search was conducted of all TV literature published from 9 January 2004 through 24 September 2008. Approximately 175 pertinent abstracts and articles were reviewed and discussed with national experts. This article describes advances in TV diagnostics which have led to an improved understanding of the epidemiology of this pathogen, as well as potential biologic and epidemiological interactions between TV and human immunodeficiency virus (HIV). New data on treatment outcomes, metronidazole-resistant TV, management of nitroimidazole-allergic patients, frequency of recurrent TV infection following treatment, and screening considerations for TV in certain populations are also presented.

Background Detection of semen biomarkers in vaginal fluid can be used to assess women's recent exposure to semen. Quantitative tests for detection of prostate-specific antigen (PSA) perform well, but are expensive and require specialized equipment. We assessed two rapid immunochromatographic strip tests for identification of semen in vaginal swabs. Study Design We tested 581 vaginal swabs collected from 492 women. Vaginal secretions were eluted into saline, and PSA was measured using the quantitative IMx (Abbott Laboratories, Abbott Park, IL, USA) assay. Specimens were also tested using the ABAcard p30 test (Abacus Diagnostics, West Hills, CA, USA) for detection of PSA and RSID-Semen test (Independent Forensics, Hillside, IL, USA) for detection of semenogelin (Sg). Results Vaginal swab extraction using saline was compatible with direct assessment of vaginal swab eluates using ABAcard for PSA detection, but not for Sg detection using RSID. The rapid PSA test detected 91% of specimens containing semen compared to 74% by the rapid Sg test. Conclusion Investigators are urged to optimize vaginal swab specimen preparation methods for performance of RSID or other tests to detect semen components other than PSA. Previously described methods for PSA testing are not uniformly applicable to other tests.

HIV-1 is present in anatomical compartments and bodily fluids. Most transmissions occur through sexual acts, making virus in semen the proximal source in male donors. We find three distinct relationships in comparing viral RNA populations between blood and semen in men with chronic HIV-1 infection, and we propose that the viral populations in semen arise by multiple mechanisms including: direct import of virus, oligoclonal amplification within the seminal tract, or compartmentalization. In addition, we find significant enrichment of six out of nineteen cytokines and chemokines in semen of both HIV-infected and uninfected men, and another seven further enriched in infected individuals. The enrichment of cytokines involved in innate immunity in the seminal tract, complemented with chemokines in infected men, creates an environment conducive to T cell activation and viral replication. These studies define different relationships between virus in blood and semen that can significantly alter the composition of the viral population at the source that is most proximal to the transmitted virus., Author Summary The work described in this report is directed at how HIV-1 viral RNA populations differ between the blood plasma and male genital tract in established infection. This site is of special interest since it is the proximal source of most transmissions of HIV-1. Thus, lessons learned about HIV-1 in the seminal tract are directly relevant to the mechanism of HIV-1 transmission. We have used single genome amplification to generate viral sequences from paired blood and semen samples in men with chronic HIV-1 infection. When compared to viral populations in blood plasma, we observe that virus in the seminal plasma can be equilibrated, clonally-amplified, or compartmentalized. We have also performed a characterization of the cytokine and chemokine milieu in these two compartments. We report a dramatic concentration of immune modulators in the seminal plasma relative to the blood, and these likely enhance the potential for viral replication in this compartment by creating an environment where target cells are kept in an activated state. These data define new and distinct features of virus:host interactions and represent a significant advance in our understanding of HIV-1 replication in the male genital tract.

Challenges in the accurate measurement of sexual behavior in human immunodeficiency virus (HIV) prevention research are well documented and have prompted discussion about whether valid assessments are possible. Audio computer-assisted self-interviewing (ACASI) may increase the validity of self-reported behavioral data. In 2006–2007, Zimbabwean women participated in a randomized, cross-sectional study that compared self-reports of recent vaginal sex and condom use collected through ACASI or face-to-face interviewing (FTFI) with a validated objective biomarker of recent semen exposure (prostate-specific antigen (PSA) levels). Of 910 study participants, 196 (21.5%) tested positive for PSA, an indication of semen exposure during the previous 2 days. Of these 196 participants, 23 (11.7%) reported no sex in the previous 2 days, with no difference in reported sexual activity between interview modes (12.5% ACASI vs. 10.9% FTFI; Fisher's exact test: P = 0.72). In addition, 71 PSA-positive participants (36.2%) reported condom-protected vaginal sex only; their reports also indicated no difference between interview modes (33.7% ACASI vs. 39.1% FTFI; P = 0.26). Only 52% of PSA-positive participants reported unprotected sex during the previous 2 days. Self-report was a poor predictor of recent sexual activity and condom use in this study, regardless of interview mode, providing evidence that such data should be interpreted cautiously.

Research evaluating clinical or behavioral interventions, barrier methods, or microbicides for the prevention of HIV and other sexually transmitted infections frequently relies on self-reports of sexual behavior, which may be inaccurate. The validity of self-reported data relating to coitus is threatened by social desirability bias, leading to concerns that study subjects may frequently fail to report unprotected sex.1 The detection of seminal biomarkers in vaginal fluid provides objective evidence of a woman’s recent exposure to semen. Results from recent studies comparing detection of prostate-specific antigen (PSA) in vaginal swabs with self-reported condom use among female sex workers suggest that concerns about misreported semen exposure may be well-founded; substantial proportions of women who reported no sex or protected sex only within the past 48 hours, tested positive for PSA.2,3 PSA (also known as p30) has been validated as a reliable marker of semen exposure in studies of vaginal specimens obtained after unprotected intercourse, or after vaginal insemination with different volumes of semen.4–6 High PSA concentrations (100–10000 ng PSA/mL vaginal swab eluate) are detectable immediately after exposure, and levels return to baseline (

BACKGROUND:: Hormonal contraception use by women may increase the risk of acquiring certain sexually transmitted infections. We explored the effect of hormonal contraceptive use specifically oral contraception (OC) and depot-medroxyprogesterone acetate (DMPA) on Trichomonas vaginalis infections in women. METHODS:: We examined data from a prospective case-control study of women with trichomoniasis and noninfected female patients recruited from 3 public sexually transmitted disease clinics. Women with positive wet mount microscopy or T. vaginalis culture results were classified as having trichomoniasis. Participants underwent physical examinations sexually transmitted infections testing and completed questionnaires which included information about demographics sexual behavior douching and contraceptive use. We assessed the association between hormonal contraceptives and trichomoniasis using bivariable and multivariable analysis and estimated exposure odds ratios (ORs) and adjusted odds ratios (aORs) with 95% confidence intervals (CIs). RESULTS:: We identified 427 women with trichomoniasis and 144 uninfected women who had information reported about contraception use. Compared with nonhormonal contraceptive use OC use was negatively associated with trichomoniasis in bivariable analysis (OR: 0.5; 95% CI: 0.3-0.8). This association was no longer statistically significant after adjusting for demographic variables douching and condom use (aOR: 0.9; 95% CI: 0.5-1.6). Use of DMPA compared with nonhormonal contraceptive use was not associated with trichomoniasis in bivariable or multivariable analyses (OR: 1.0 95% CI: 0.5-2.1; aOR = 1.4 95% CI: 0.6-3.4 respectively). CONCLUSIONS:: Although OC use appeared to have a protective effect in the bivariable analysis the hormonal contraceptives OC and DMPA were not associated with T. vaginalis infection after adjustment for other factors.

The clinical significance of Mycoplasma genitalium (MG) infection in adolescent women is poorly understood. We compared the prevalence of MG with that of other sexually transmitted organisms such as Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), and Trichomonas vaginalis (TV) and assessed the associations of MG with sexual behaviors, genitourinary symptoms, physical and laboratory findings.Women aged 14 to 21 years (n = 331) were recruited from an urban medical center. The subjects' sexual behaviors, genitourinary symptoms, and physical findings were recorded. Endocervical swabs were collected for CT and NG testing and vaginal swabs for wet mount, Gram stain, TV and MG testing. MG infection was identified by nucleic acid amplification using a transcription-mediated amplification assay.MG was detected in 74 (22.4%), CT in 79 (24.4%), TV in 60 (18.2%), and NG in 35 (10.7%) subjects. MG infection was not associated with vaginal symptoms, physical evidence of cervicitis, or findings on wet mount or Gram stain. In logistic regression, variables positively associated with MG were current CT [odds ratio (OR), 2.3; 95% confidence interval (CI), 1.4-4.4] and recent sexual contact (or =7 days) (OR, 2.0; CI, 1.1-3.2). Dysuria (OR, 0.44; CI, 0.2-0.96) and use of hormonal contraception (OR, 0.55; CI, 0.3-1.0) were negatively associated with MG infection.In adolescent women, MG infection was as common as chlamydial infection and trichomoniasis and more common than gonorrhea. MG was associated with CT and recent sexual contact but not with vaginal symptoms or signs of cervicitis.

At the beginning of the 21st century, we face stable or increasing infection rates for Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (GC) in the United States. These are unacceptable circumstances in view of the serious negative impact of these infections on reproductive health and their relatively high prevalence in the United States compared with other industrialized countries.1,2 Clearly, reduction of prevalence and incidence requires improved intervention approaches, including better screening strategies. CT and GC infections in the general population are often asymptomatic and consequently are underdiagnosed.3,4 Targeted screening programs in the United States have resulted in steady declines in the prevalence of these infections in the 1990s, a trend that unfortunately has not been sustained in recent years. Surveillance statistics from the CDC reveal that reported cases of gonorrhea rose sharply from the mid 1960s through the 1970s, declined steadily until the mid 1990s and then plateaued.2 Comparable data for CT infections have been collected only since 1984, and nationwide, case rates appear to have risen steadily through the most recent annual data summary period in 2005. However, screening for CT became widespread only after the advent of affordable, reliable detection methods including nucleic acid amplification tests (NAATs) in the 1990s, and case detection has been increasing steadily ever since.2 Thus, the apparent increase in CT case detection is likely due in part to increased case finding. However, in the northwestern states comprising Public Health Region X, funding for broad-based CT screening among women was initiated in 1988, and data have been collected on the number of tests performed and the percent of CT tests that were positive. In Region X, CT prevalence rates actually dropped steadily between 1988 and 1995 and then remained steady through 1997 (Fig. 1).5 Data from the regional Infertility Prevention Programs have provided good CT prevalence data from participating family planning clinics in the United States since the late 1990s. Infertility Prevention Programs data show that CT rates in women seen in these settings remained steady nationwide through 2005.5 Importantly, in Region X, where screening programs have been in place the longest, the percentage of women tested who have positive tests for CT is now rising, even after statistical adjustment for increases in test sensitivity (Fig. 1). The reasons for the lack of continued decline (and even increases) in both CT and GC prevalence are not entirely clear, but these trends may be consequences of initial screening programs that targeted only a subset of the population (those who attended the clinics), allowing continued spread in underserved sectors of the community (those who did not attend the clinics). Thus, it is logical to conclude that programs for screening all at-risk populations could further reduce sexually transmitted infection (STI) prevalence. Fig. 1 Trends in Chlamydia trachomatis positivity among 15- to 24-year old women tested in family planning clinics in northwestern states of Public Health Region X, 1988 to 2005. Data are from the Centers for Disease Control and Prevention.5 Black bars are unadjusted, ... Possible strategies for reducing STI prevalence include expanding screening to nontraditional settings such as schools, juvenile detention centers, hospital emergency departments, and drug rehabilitation clinics. The availability of highly sensitive NAATs allows the use of specimens other than conventional clinician-obtained endocervical and urethral specimens, such as urine and vaginal specimens, for detection of CT and GC.

The objectives were to evaluate the association between self-reported condom use and prevalent and incident chlamydia gonorrhea and trichomoniasis. Prospective study of 414 males attending a sexually transmitted infection (STI) clinic in Jamaica. Condom use and STI status were assessed at enrollment and at 4 follow-up visits. The analyses on condom use and prevalent STI included data from 414 men while those on incident STI were based on 1111 intervals from 355 men. We diagnosed prevalent STI (chlamydia gonorrhea and/or trichomoniasis) in 54.6% (n = 226) of the participants at enrollment. About 14% (n = 51) of participants had at least 1 of the study STIs during follow-up. Follow-up visits in which participants reported consistent condom use (100% of acts) for the past 7 days had less incident STI (adjusted OR 0.4; 95% CI 0.2-0.9) compared with visits where no condom use was reported. Self-reported condom use was more closely correlated with incident than prevalent STI. For example the adjusted OR for prevalent infection for participants reporting consistent versus no condom use in past 7 days was 0.7 (95% CI 0.4-1.2). Classifications based on the number of unprotected acts yielded findings similar to those based on the proportion of acts protected. Consistent condom use was associated with reduced risk of incident urethral STI. Research on condom effectiveness should focus on incident STI outcomes where the temporal relationship between condom use and infection is clearer. (authors)

The effect that vaginal lactobacilli have on trichomoniasis is not known. At 3 clinics for treatment of sexually transmitted disease, we recruited 521 female patients with trichomoniasis and 176 control subjects. All participants underwent physical examinations and testing for sexually transmitted infection and completed questionnaires. Low levels of vaginal lactobacilli were associated with trichomoniasis (odds ratio [OR], 2.2 [95% confidence interval {CI}, 1.4-3.2]). After adjustment for covariables, this association remained in women with a higher educational level (OR, 4.6 [95% CI, 2.2-9.5]) but not in women with a lower educational level (OR, 1.6 [95% CI, 0.7-3.4]). Vaginal lactobacilli may be associated with trichomoniasis in women with higher levels of education or a related behavioral factor.

Trichomonas vaginalis infection in men is an important cause of nongonococcal urethritis. Effective detection of the parasite in men using culture requires examination of multiple specimens. We compared culture and PCR-enzyme-linked immunosorbent assay in urethral swabs, urine, and semen for T. vaginalis detection in male sexual partners of women with trichomoniasis identified by wet mount and culture. Trichomonads were detected by at least one positive test in 205/280 men (73.2%) who submitted at least one specimen for culture and PCR. Whereas InPouch TV culture detected only 46/205 cases (22.5%), PCR detected 201/205 (98.0%). Urethral swab cultures from men with urethritis were more likely to be positive with shorter incubation than specimens from men without urethritis. T. vaginalis was detected more often in men with wet-mount-positive partners. Even with a sensitive PCR assay, reliable detection of T. vaginalis in male partners required multiple specimens. The majority of male sexual partners in this study were infected, emphasizing the importance of partner evaluation and treatment.

The etiologic agent of chancroid isHaemophilus ducreyi. To fulfill its obligate requirement for heme,H. ducreyiuses two TonB-dependent receptors: the hemoglobin receptor (HgbA) and a receptor for free heme (TdhA). Expression of HgbA is necessary forH. ducreyito survive and initiate disease in a human model of chancroid. In this study, we used a swine model ofH. ducreyiinfection to demonstrate that an experimental HgbA vaccine efficiently prevents chancroid, as determined by several parameters. Histological sections of immunized animals lacked typical microscopic features of chancroid. All inoculated sites from mock-immunized pigs yielded viableH. ducreyicells, whereas no viableH. ducreyicells were recovered from inoculated sites of HgbA-immunized pigs. Antibodies from sera of HgbA-immunized animals bound to and initiated antibody-dependent bactericidal activity against homologousH. ducreyistrain 35000HP and heterologous strain CIP542 ATCC; however, an isogenichgbAmutant of 35000HP was not killed, proving specificity. Anti-HgbA immunoglobulin G blocked hemoglobin binding to the HgbA receptor, suggesting a novel mechanism of protection through the limitation of heme/iron acquisition byH. ducreyi. Such a vaccine strategy might be applied to other bacterial pathogens with strict heme/iron requirements. Taken together, these data suggest continuing the development of an HgbA subunit vaccine to prevent chancroid.

Background: Too little is known about the many women who generate income in Madagascar by trading sex. Methods: Clinical and laboratory exams were offered to 493 non-care seeking female sex workers (SWs) in Antananarivo and 493 in Tamatave. SWs were recruited by peers in their community; they were interviewed, counselled and treated for sexually transmitted infections (STIs) at recruitment and re-evaluated 2 months later. Results: One hundred and eighty six (38%) of the SWs in Antananarivo and 113 (23%) in Tamatave did not complete primary school (P < 0.0001). The incidence rates per person per month in Antananarivo and Tamatave, respectively, were 0.09 and 0.08 for gonorrhoea; 0.05 and 0.03 for chlamydia; 0.24 and 0.15 for trichomoniasis; 0.07 and 0.05 for syphilis. At follow-up, consistent condom use with clients was reported by 56 (12%) SWs in Antananarivo and 137 (29%) in Tamatave (P < 0.0001); 320 (70%) SWs in Antananarivo and 11 (2%) in Tamatave reported sex with a non-paying partner in the past month (P < 0.0001). In Antananarivo, 422 (92%) of the SWs thought they were at no or low risk of having an STI compared to 100 (21%) in Tamatave (P = 0.02). At follow-up, 277 (61%) SWs reported no birth control for their last sex act in Antananarivo, compared to 26 (5%) in Tamatave (P < 0.0001). Socio-demographic and behavioural risk factors for incident gonorrhoea, chlamydia and trichomoniasis varied by city. Conclusions: Strategies to address the needs of women who trade sex should include educational and economic opportunities; should tackle male partners of SWs; promote dual protection against unintended pregnancy and STIs, while taking into account local socio-demographic and behavioural characteristics.

Background and Objectives: The prevalence of trichomoniasis in the general population of the United States is unknown. This study provides the first population-based prevalence estimates of trichomoniasis among young adults in the United States. Methods: The National Longitudinal Study of Adolescent Health (Add Health) is an ongoing prospective cohort study. In a cross-sectional analysis of Wave III of Add Health (N = 12,449), we determined the prevalence of trichomoniasis using a polymerase chain reaction assay. Results: The estimated overall prevalence of trichomoniasis in U.S. young adults was 2.3% (95% confidence interval [CI], 1.8–2.7%). The prevalence was slightly higher among women (2.8%; 95% CI, 2.2–3.6%) than men (1.7%; 95% CI, 1.3–2.2%). The prevalence increased with age and varied by region, with the south having the highest prevalence (2.8%; 95% CI, 2.2–3.5%). The prevalence was highest among black women (10.5%; 95% CI, 8.3–13.3%) and lowest among white women (1.1%; 95% CI, 0.8–1.6%). Among men, the prevalence was highest among Native Americans (4.1%; 95% CI, 0.4–29.3%) and blacks (3.3%; 95% CI, 2.2–4.9%), and lowest among white men (1.3%; 95% CI, 0.9–1.8%). Conclusions: Trichomoniasis is moderately prevalent among the general U.S. population of young adults and disturbingly high among certain racial/ethnic groups.

Context: Chlamydial and gonococcal infections are important causes of pelvic inflammatory disease, ectopic pregnancy, and infertility. Although screening for Chlamydia trachomatis is widely recommended among young adult women, little information is available regarding the prevalence of chlamydial and gonococcal infections in the general young adult population. Objective: To determine the prevalence of chlamydial and gonoccoccal infections in a nationally representative sample of young adults living in the United States. Design, Setting, and Participants: Cross-sectional analyses of a prospective cohort study of a nationally representative sample of 14322 young adults aged 18 to 26 years. In-home interviews were conducted across the United States for Wave III of The National Longitudinal Study of Adolescent Health (Add Health) from April 2, 2001, to May 9, 2002. This study sample represented 66.3% of the original 18924 participants in Wave I of Add Health. First-void urine specimens using ligase chain reaction assay were available for 12548 (87.6%) of the Wave III participants. Main Outcome Measures: Prevalences of chlamydial and gonococcal infections in the general young adult population, and by age, self-reported race/ethnicity, and geographic region of current residence. Results: Overall prevalence of chlamydial infection was 4.19% (95% confidence interval [CI], 3.48%-4.90%). Women (4.74%; 95% CI, 3.93%-5.71%) were more likely to be infected than men (3.67%; 95% CI, 2.93%-4.58%; prevalence ratio, 1.29; 95% CI, 1.03-1.63). The prevalence of chlamydial infection was highest among black women (13.95%; 95% CI, 11.25%-17.18%) and black men (11.12%; 95% CI, 8.51%- 14.42%); lowest prevalences were among Asian men (1.14%; 95% CI, 0.40%-3.21%), white men (1.38%; 95% CI, 0.93%-2.03%), and white women (2.52%; 95% CI, 1.90%-3.34%). Prevalence of chlamydial infection was highest in the south (5.39%; 95% CI, 4.24%-6.83%) and lowest in the northeast (2.39%; 95% CI, 1.56%-3.65%). Overall prevalence of gonorrhea was 0.43% (95% CI, 0.29%-0.63%). Among black men and women, the prevalence was 2.13% (95% CI, 1.46%-3.10%) and among white young adults, 0.10% (95% CI, 0.03%-0.27%). Prevalence of coinfection with both chlamydial and gonococcal infections was 0.030% (95% CI, 0.18%-0.49%). Conclusions: The prevalence of chlamydial infection is high among young adults in the United States. Substantial racial/ethnic disparities are present in the prevalence of both chlamydial and gonococcal infections.

In this study, we examined the immune response during gonococcal infection to the individual transferrin binding proteins by using a quantitative enzyme-linked immunosorbent assay (ELISA). Recombinant transferrin binding protein A (rTbpA) and rTbpB were purified under nondenaturing conditions for use as ELISA antigens. Sera and secretions from culture-positive individuals were analyzed for antibodies to rTbpA and rTbpB and compared to samples from individuals with no history of gonococcal infection. Although antibodies to both rTbpA and rTbpB were detected in serum, in most cases the antibody levels were not significantly different from those measured in the control population. Also, previous history of gonococcal infection did not increase antibody levels in serum, suggesting the lack of an anamnestic response. Analysis of secretion samples revealed antibody levels that were generally below the limits of detection in our assay. Overall, this study demonstrated a paucity of systemic and local antibody responses to rTbps as a result of natural infection and represents a baseline over which a protective antibody response will have to be generated in order to develop an efficacious gonococcal vaccine.

All isolates of Neisseria gonorrhoeae express receptors that bind human transferrin (Tf). Although lactoferrin (Lf) is abundant on mucosa and in purulent exudates, many gonococci do not express an Lf receptor. The naturally occurring Lf receptor deletion mutant FA1090 (LbpB-LbpA-) is infectious, but a Tf receptor mutant of FA1090 is unable to infect male volunteers [Cornelissen, C.N., Kelley, M., Hobbs, M.M., Anderson, J.E., Cannon, J.G., Cohen, M.S., and Sparling, P.F. (1998) Mol Microbiol 27: 611-616]. Here, we report that expression of an Lf receptor in the absence of the Tf receptor was sufficient for infection, and that expression of both Lf and Tf receptors resulted in a competitive advantage over a strain that made only the Tf receptor in mixed infection of male volunteers. We confirmed that nearly 50% of clinical isolates do not make an Lf receptor. Surprisingly, about half of geographically diverse Lf - isolates representing many different auxotypes and porin serovars carried an identical lbpB lbpA deletion. Among Lf+ strains, all produced the integral outer membrane protein LbpA, but 70% did not express the lipoprotein LbpB. Thus, there are apparently selective pressures for expression of the Lf receptor in the male urethra that are balanced by others against expression of the Lf receptor in niches other than the male urethra.

Trichomonas vaginalis infection is highly prevalent worldwide and is associated with urethritis, prostatitis, and urethral strictures in men. However, the natural history and importance of T. vaginalis in men are poorly understood, in part because of difficulties in diagnosing infection. Traditional detection methods rely on culture and wet-mount microscopy, which can be insensitive and time consuming. Urethral swabs are commonly used to detect T. vaginalis in men, but discomfort from specimen collection is a barrier to large studies. One thousand two hundred twenty-five Malawian men attending sexually transmitted disease and dermatology clinics were enrolled in this cross-sectional study to validate detection by urine-based PCR-enzyme-linked immunosorbent assay (ELISA) with urine and urethral swab culture as the reference standard. This assay for detection of amplified T. vaginalis DNA in first-catch urine (≤30 ml) performed with a sensitivity of 92.7%, a specificity of 88.6%, and an adjusted specificity of 95.2% compared to culture of urethral swabs or urine sediment. For clinical research settings in which urethral swabs are not available and culture is not feasible, the urine-based PCR-ELISA may be useful for detection of trichomoniasis in men.

PenB is the third resistance determinant in the stepwise acquisition of multiple resistance genes in chromosomally mediated resistant Neisseria gonorrhoeae (CMRNG). Alterations in por IB , one of two alleles at the por locus that encodes the outer membrane protein porin IB (PIB), were recently reported to be responsible for the increased resistance to penicillin and tetracycline conferred by penB , but the specific mutations conferring antibiotic resistance were not identified experimentally. To determine which amino acids in PIB confer increased resistance, we transformed a recipient strain with chimeras of the por IB genes from strains FA1090 and FA140 ( penB2 ). These studies revealed that two amino acid changes, G120D and A121D, were both necessary and sufficient to confer increased resistance to penicillin and tetracycline. Site-saturation and site-directed mutagenesis of Gly-120 and Ala-121 revealed that both a single mutation, G120K, and the double mutations G120R A121H and G120P A121P also conferred antibiotic resistance to the recipient strain. The identical mutations in PIA increased penicillin and tetracycline resistance either moderately or not at all. Analysis of por IB genes present in the GenBank database from 51 clinical isolates demonstrated that lysine and aspartate mutations at positions 120 and/or 121 also occur in nature. These studies demonstrate that charged amino acids at positions 120 and 121 in PIB are highly preferential for conferring resistance to penicillin and tetracycline in N. gonorrhoeae .

Trichomonas vaginalis infection is highly prevalent worldwide and is associated with poor birth outcomes and enhanced human immunodeficiency virus transmission. Traditional detection methods rely on microscopic examination of vaginal specimens (wet mount) and culture, which can be insensitive and time-consuming. More than 3,000 women attending two sexually transmitted disease clinics were enrolled in this cross-sectional study to evaluate urine-based PCR for detection of T. vaginalis using a combined reference standard of wet mount and culture from vaginal swab. The prevalence of trichomoniasis in the population was 16.7% (502 of 3,009 women) using the reference standard. PCR with urine combined with agarose gel-based detection was 66.9% sensitive and 98.3% specific compared to the reference standard. Detection of PCR products using an unlabeled enzyme-linked immunosorbent assay (ELISA) improved the sensitivity to 86.4%, but specificity fell to 86.1%. Using a digoxigenin-labeled ELISA for detection of amplified T. vaginalis DNA from urine, the sensitivity and specificity of the PCR improved to 90.8 and 93.4%, respectively, compared to wet mount or culture from vaginal swabs. For clinical research settings in which vaginal specimens are not available and culture conditions are not feasible, urine-based PCR-ELISA may be useful for the detection of trichomoniasis in women.

Background In 2007, 201 men who have sex with men were recruited at social venues and through peer referral in five urban areas of Jamaica for an exploratory study of the prevalence of sexually transmitted infection in the population and identification of risk factors. Objective To describe the association between age and current infection with HIV, syphilis, and chlamydia and explore reasons for the differences. Methods MSM were invited to participate in a survey and testing. Urine and blood samples were collected at the time of the interview and later tested for evidence of HIV, syphilis, gonorrhoea and chlamydia. Men were given options about where to obtain results. Results Of the three infections, HIV had the strongest positive association with age; syphilis a weaker positive association; and chlamydia a weak negative association. Risk factors varied somewhat by age and infection see Abstract P1-S2.63 Figure 1. For example, not using a condom in the past 4 weeks was associated with chlamydia infection among MSM age 15−19 but not for older MSM. Having a main sexual partner was associated with HIV infection for MSM age 15−19 but not for older MSM. Conclusion Strategies for prevention of transmission need to be aware of differences in risk by age and by infection.

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 for Neisseria gonorrhoeae (NG) and Chlamydia trachomatis (CT) using APTIMA Combo2 and for Trichomonas 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.

Background This study evaluated the clinical performance of the APTIMA® Trichomonas vaginalis (ATV, Gen-Probe Incorporated) Assay, a nucleic acid amplification test for the diagnosis of Trichomonas vaginalis (TV) infection, in asymptomatic and symptomatic women. Methods This prospective, multicenter clinical trial enrolled 1025 women attending US OB-GYN, adolescent, family planning, or sexually transmitted disease clinics. Four specimen types were collected from each subject: physician-collected vaginal swab, endocervical swab, ThinPrep specimen, and first-catch urine. Of three vaginal swabs collected from each subject, one was used for wet mount microscopic examination, one for culture, and one for molecular testing for TV. The order of collection for each vaginal swab sample was rotated to minimise sampling bias. Each specimen was tested by ATV assay using the automated TIGRIS DTS system. ATV assay performance in each sample type was determined by comparing ATV assay test result to the patient infected status (positive in saline wet mount and/or culture) for each sample. Results Of 933 subjects included in the analysis, 59.9% were symptomatic. Compared to patient infected status, ATV assay clinical sensitivities and specificities were 100% and 99.0%, respectively, from vaginal swabs; 100% and 99.4%, respectively, from endocervical swab; 100% and 99.6%, respectively, from ThinPrep specimens; and 95.2% and 98.9%, respectively, from patient-collected urine samples. ATV assay performance was similar in asymptomatic and symptomatic patients, by age group (14–17 years and 18 years or older), and was consistent between testing sites. The ATV assay also demonstrated superior performance compared to wet mount microscopic examination and TV culture, regardless of the specimen type utilised. Conclusions This study provides clinical validation of the ATV assay for the intended uses of detecting TV rRNA in asymptomatic women, and/or to aid in the diagnosis of trichomoniasis in symptomatic women, in a US population. The use of highly accurate, fully-automated molecular tests such as the ATV assay for testing easily obtained vaginal swab and urine samples should facilitate large-scale screening for TV in the US.

Objectives To assess the potential impact of chlamydial screening policy that recommends routine screening of women but not men. Methods Population surveys of probability samples of Baltimore adults aged 18 to 35 years in 1997–1998 and 2006–2009 collected biospecimens to estimate trends in undiagnosed chlamydial infection. Survey estimates are compared to surveillance data on diagnosed chlamydial infections reported to the Health Department. Results Prevalence of undiagnosed chlamydial infection among men increased from 1.6% to 4.0%, but it declined from 4.3% to 3.1% among women (p = 0.028 for test of interaction). The annual (average) number of diagnosed infections was substantially higher among women than men in both time periods and increased among both men and women. Undiagnosed infection prevalence was substantially higher among black than non-black adults (4.0% vs 1.2%, p = 0.042 in 1997–98 and 5.5% vs 0.7%, p

All gonococci encode a hemoglobin (Hgb) receptor, but it is phase variable, and most laboratory and clinical isolates are in the Hgb receptor "off" phase. In the present study, we address the question of whether there is a selective advantage to expressing the Hgb receptor during early phases of the menstrual cycle, when Hgb is readily available from menstrual blood. Inclinical isolates collected from women, Hgb use in vitro (Hgb receptor "on" phase) was associated with a shorter time since the onset of the last menstrual cycle, (P=.031, Wilcoxon rank-sum test). Thus, there may be a selective advantage to expression of the gonococcal Hgb receptor during infection of women in the early phases of their menstrual cycles.

Thirty-three Neisseria gonorrhoeae isolates from 15 persons infected multiple times with the same serovar were compared using por gene sequencing, opa-typing, and arbitrarily primed-polymerase chain reaction. All three molecular techniques were more discriminatory than serotyping and identified differences between some isolates belonging to the same serovar. Although there were differences among Por sequences within some serovars, 10 of 15 subjects became reinfected with gonococci expressing identical Por proteins. Sequence analysis of por genes revealed evidence of horizontal genetic exchange and point mutations in potential surface-exposed regions during passage in the community.

Iron, an essential nutrient for most microorganisms, is sequestered by the host to decrease the concentration of iron available to bacterial pathogens. Neisseria gonorrhoeae, the causative agent of gonorrhoea, can acquire iron by direct interaction with human iron-binding proteins, including the serum glycoprotein, transferrin. Iron internalization from host transferrin requires the expression of a bacterial receptor, which specifically recognizes the human form of transferrin. Two gonococcal transferrin-binding proteins have been implicated in transferrin receptor function, TbpA and TbpB. We constructed a gonococcal transferrin receptor mutant without the introduction of additional antibiotic resistance markers and tested its ability to cause experimental urethritis in human male volunteers. The transferrin receptor mutant was incapable of initiating urethritis, although the same inoculum size of the wild-type parent strain, FA1090, causes urethritis in >90% of inoculated volunteers. To our knowledge, this is the first experimental demonstration that a bacterial iron acquisition system is an essential virulence factor for human infection.

Haemophilus ducreyi causes chancroid, a sexually transmitted genital ulcer disease implicated in increased heterosexual transmission of HIV. As part of an effort to identify H. ducreyi gene products involved in virulence and pathogenesis, we created random TnphoA insertion mutations in an H. ducreyi 35000 library cloned in Escherichia coli. Inserts encoding exported or secreted PhoA fusion proteins were characterized by DNA sequencing. One such clone encoded a Cu-Zn superoxide dismutase (SOD) enzyme. The Cu-Zn SOD was periplasmic in H. ducreyi and accounted for most of the detectable SOD activity in whole-cell lysates of H. ducreyi grown in vitro. To investigate the function of the Cu-Zn SOD, we created a Cu-Zn SOD-deficient H. ducreyi strain by inserting a cat cassette into the sodC gene. The wild-type and Cu-Zn SOD null mutant strains were equally resistant to excess cytoplasmic superoxide induced by paraquat, demonstrating that the Cu-Zn SOD did not function in the detoxification of cytoplasmic superoxide. However, the Cu-Zn SOD null strain was significantly more susceptible to killing by extracellular superoxide than the wild type. This result suggests that the H. ducreyi Cu-Zn SOD may play a role in bacterial defence against oxidative killing by host immune cells during infection.

Summary: Opacity (Opa) proteins are a family of antigenically variable outer-membrane proteins of Neisseria meningitidis. ET-37 complex meningococci, defined by multilocus enzyme electrophoresis, have been isolated on different continents. Twenty-six different Opa proteins have been observed within strains of the ET-37 complex isolated between the 1960s and the 1980s, although individual strains have only four opa genes per chromosome. In this work the opa genes of four closely related ET-37 complex N. meningitidis strains recently isolated from Mali, West Africa were characterized and compared with the opa genes of strain FAM18, an ET-37 complex isolate from the USA. DNA sequence analysis and Southern blot experiments indicated that recombinational reassortment, including gene duplication and import by horizontal genetic exchange, has occurred in the opa genes within the ET-37 complex, resulting in two partially different Opa repertoires being present in FAM18 and the Mali isolates. Using synthetic peptides derived from the hypervariable (HV) regions of opa genes, the epitopes for nine mAbs were mapped. These bacteria, isolated on different continents, contain both shared and unique opa HV regions encoding epitopes recognized by mAbs and show evidence of recombinational reassortment of the HV regions.

Background The availability of sensitive point-of-care tests for Trichomonas vaginalis (TV) in emergency rooms (ER) is important to facilitate immediate diagnosis and treatment. We evaluated the use of the OSOM ® Trichomonas Rapid Test, an FDA-approved immunochromatographic capillary flow assay, among female patients in an ER located in the southeastern United States. Methods The University of North Carolina Hospitals located in Chapel Hill, North Carolina, US replaced wet mount microscopy (WM) for vaginal TV detection with the OSOM ® Trichomonas Rapid Test (Genzyme Corporation, US) in October 2011. We analysed 10 months of data for women evaluated in the ER with the rapid test, and compared the positivity rate with a similar 10-month period determined by WM. We assessed characteristics of women identified with trichomoniasis using the rapid test, and the proportion who received appropriate therapy. Results Over the study period, 1533 female patients were tested for TV using the rapid test, of which 539 (35.2%) were Black, with a median age of 28 years (range 6–82 years). The TV positivity rate based on the rapid test was 5.6% (86/1533), compared to 3.2% (54/1681) among those tested with WM. Among 77 unique female patients with TV infection, 467 (34.0%) were Black and had a median age of 28 years. Nearly half of infected women presented with a chief complaint of abdominal pain (49%); vaginal discharge was only reported by 10%. Among infected women, 55% had concomitant bacterial vaginosis (BV), and 16% were co-infected with gonorrhoea and/or chlamydia. Most infected women (84%) were prescribed metronidazole during the same ER visit. Conclusions . The OSOM ® Trichomonas Rapid Test resulted in a 60% increase in TV detection among women compared to WM, and the majority received appropriate TV therapy. Women identified with TV infection in an ER setting were primarily co-infected with BV and other STIs.