Low-cost conventional PCR techniques enable simultaneous detection of bacterial sexually transmitted infections with enhanced sensitivity and specificity.

PURPOSE: Neisseria gonorrhoeae (NG), Chlamydia trachomatis (CT) and Mycoplasma hominis (MH), the three most common treatable bacterial sexually transmitted infections (STIs) worldwide can lead to many complications if remain untreated. Screening of high-risk population with highly sensitive methods will lead to significant improvement in patient outcomes and will prevent downward transmission. The advantages of Polymerase chain reaction (PCR) based assay are not only high sensitivity and specificity, but also detection of multiple organisms in a single reaction which reduce the result turn-around time. The aim of the present study was to evaluate the feasibility of a multiplex PCR assay method targeting 16S rRNA gene for simultaneous detection of NG, CT and MH infection along with their trend and occurrence among high-risk population in Assam, Northeast India. METHODS: A cross-sectional study was undertaken, where a total of 200 randomly selected patients from high-risk population were included. After validation of singleplex PCR, Multiplex PCR (M-PCR) was performed along with the traditional culture method for NG. RESULTS & CONCLUSION: The overall agreement of M-PCR with singleplex PCR was very high (100%). The occurrence of STI was found to be very high (101/200; 50.5%). Furthermore, co-infection was detected in 10/200; 5%) individuals. Infection was more common among young individuals (p < 0.05) and males out-numbered females (p < 0.05). The most common organism detected was CT (42/200; 21%) followed by NG (41/200; 20.5%) and MH (20/200; 10%). The M-PCR assay workflow is simple, cost effective and can be used in routine diagnostic laboratories with basic molecular facilities.

Gardnerella vaginalis in urinary tract infections, are men spared?

Gardnerella vaginalis in association with anaerobes has been linked to bacterial vaginosis in women, while urinary tract infections (UTIs) in men have rarely been reported. The aim of the review was to reveal the significance of G. vaginalis UTIs in men. Prevalence of G. vaginalis UTIs in men varied from 0.5 to >27% according to patients' groups. Most patients had comorbidity such as urolithiasis or stents, transplants, tumors and diabetes, however, infections can also affect immunocompetent patients. We observed G. vaginalis-associated bacteriuria and leukocyturia in a kidney transplant man. Complications of the UTIs such as bacteremia (in 9/11 cases), hydronephrosis (4/11) and abscesses or septic emboli have been reported. Bacterial vaginosis in female partners has been a risk factor for UTIs in males. In women, biofilm Gardnerella phenotype, stabilized by Atopobium vaginae and Prevotella bivia was linked to ≥6-fold higher antibiotic resistance rates compared with the planktonic phenotype. Non-susceptibility to metronidazole and levofloxacin was found also in males. Therefore, if aerobic urine cultures are negative, urine and blood samples from male patients with predisposing factors and clinical signs of UTIs and bacteremia, can be taken. Plates should be incubated for 2-4 days in capnophilic/microaerophilic conditions, however only anaerobic incubation can help with detecting G. vaginalis strains which grow only anaerobically. Susceptibility testing of the isolates is highly important. Briefly, adherent G. vaginalis phenotype can be sexually transmissible. Despite the infrequency of G. vaginalis UTIs in men, the infections should be considered since they are often linked to severe complications.

Changes in the prevalence of self-reported sexually transmitted bacterial infections from 2010 and 2017 in two large European samples of men having sex with men-is it time to re-evaluate STI-screening as a control strategy?

BACKGROUND/OBJECTIVES: Many European countries reported increased numbers of syphilis, gonorrhoea and chlamydia diagnoses among men who have sex with men (MSM) in recent years. Behaviour changes and increased testing are thought to drive these increases. METHODS: In 2010 and 2017, two large online surveys for MSM in Europe (EMIS-2010, EMIS-2017) collected self-reported data on STI diagnoses in the previous 12 months, diagnostic procedures, STI symptoms when testing, number of sexual partners, and sexual behaviours such as condom use during the last intercourse with a non-steady partner in 46 European countries. Multivariate regression models were used to analyse factors associated with diagnoses of syphilis, gonorrhoea/chlamydia, and respective diagnoses classified as symptomatic and asymptomatic. If applicable, they included country-level screening rates. RESULTS: Questions on STI diagnoses and sexual behaviours were answered by 156,018 (2010) and 125,837 (2017) participants. Between 2010 and 2017, overall diagnoses with gonorrhoea/chlamydia and syphilis increased by 76% and 83% across countries. Increases were more pronounced for asymptomatic compared to symptomatic infections. The proportion of respondents screened and the frequency of screening grew considerably. Condomless anal intercourse with the last non-steady partner rose by 62%; self-reported partner numbers grew. Increased syphilis diagnoses were largely explained by behavioural changes (including more frequent screening). Gonorrhoea/chlamydia increases were mainly explained by more screening and a change in testing performance. A country variable representing the proportion of men screened for asymptomatic infection was positively associated with reporting symptomatic gonorrhoea/chlamydia, but not syphilis. DISCUSSION/CONCLUSION: The positive association of country-level screening rates with the proportion of symptomatic infections with gonorrhoea/chlamydia may indicate a paradoxical effect of screening on incidence of symptomatic infections. Treatment of asymptomatic men might render them more susceptible to new infections, while spontaneous clearance may result in reduced susceptibility. Before expanding screening programmes, evidence of the effects of screening and treatment is warranted.

The Impact of Selected Bacterial Sexually Transmitted Diseases on Pregnancy and Female Fertility.

Sexually transmitted infections (STIs) caused by Neisseria gonorrhoeae, Chlamydia trachomatis and Mycoplasma genitalium are a common cause of pelvic inflammatory disease (PID) which can lead to tubal factor infertility (TFI). TFI is one of the most common causes of infertility, accounting for 30% of female fertility problems. STIs can also have an impact on pregnancy, leading to adverse pregnancy outcomes. Escalating antibiotic resistance in Neisseria gonorrhoeae and Mycoplasma genitalium represents a significant problem and can be therapeutically challenging. We present a comprehensive review of the current treatment options, as well as the molecular approach to this subject. We have given special attention to molecular epidemiology, molecular diagnostics, current and new treatments, and drug resistance.

Sociodemographic and behavioural factors associated with testing for HIV and STIs in a US nationwide sample of transgender men who have sex with men.

OBJECTIVES: Transgender men who have sex with men (TMSM) represent an understudied population in relation to screening for HIV and sexually transmitted infections (STIs). We examined HIV and STI testing prevalence among TMSM along with the factors associated with testing in a diverse US nationwide sample of TMSM. METHODS: Data from a cross-sectional online convenience sample of 192 TMSM were analysed using multivariable binary logistic regression models to examine the association between sociodemographic and behavioural factors and lifetime testing for HIV, bacterial STIs and viral STIs, as well as past year testing for HIV. RESULTS: More than two-thirds of TMSM reported lifetime testing for HIV (71.4%), bacterial STIs (66.7%), and viral STIs (70.8%), and 60.9% had received HIV testing in the past year. Engaging in condomless anal sex with a casual partner whose HIV status is different or unknown and having fewer than two casual partners in the past 6 months were related to lower odds of lifetime HIV, bacterial STI, viral STI and past year HIV testing. Being younger in age was related to lower probability of testing for HIV, bacterial STIs and viral STIs. Furthermore, TMSM residing in the South were less likely to be tested for HIV and viral STIs in their lifetime, and for HIV in the past year. Finally, lower odds of lifetime testing for viral STIs was found among TMSM who reported no drug use in the past 6 months. CONCLUSIONS: These findings indicate that a notable percentage of TMSM had never tested for HIV and bacterial and viral STIs, though at rates only somewhat lower than among cisgender MSM despite similar patterns of risk behaviour. Efforts to increase HIV/STI testing among TMSM, especially among those who engage in condomless anal sex, are needed.

Disseminated melioidosis in early pregnancy - an unproven cause of foetal loss.

BACKGROUND: Melioidosis is a potentially life-threatening infection caused by the Gram-negative bacterium Burkholderia pseudomallei. Melioidosis is difficult to diagnose due to its diverse clinical manifestations, which often delays administration of appropriate antibiotic therapy. CASE PRESENTATION: Melioidosis is uncommon in pregnancy but both spontaneous abortion and neonatal melioidosis have been reported. We report a case of bacteraemic melioidosis in a young woman with a subsequent spontaneous abortion, with B. pseudomallei cultured from a high vaginal swab as well as blood. CONCLUSION: It remains unclear in this and previously reported cases as to whether the maternal melioidosis was sexually transmitted.

Development of a SimpleProbe real-Time PCR Assay for rapid detection and identification of the US novel urethrotropic clade of Neisseria meningitidis ST-11 (US_NmUC).

Urethritis, or inflammation of the urethra, is one of the most common reasons men seek clinical care. Sexually transmitted pathogens including Neisseria gonorrhoeae are responsible for over half of the symptomatic urethritis cases in U.S. men. Recently, clinics in Indianapolis, Columbus, Atlanta, and other U.S. cities began to note increasing numbers of men presenting with urethritis and Gram-negative intracellular diplococci in their urethral smears who test negative for N. gonorrhoeae. Many of these discordant cases, which have periodically reached highs of more than 25% of presumed gonococcal cases in some sexually transmitted infection clinics in the U.S. Midwest, are infected with strains in a novel urethrotropic clade of Neisseria meningitidis ST-11 (US_NmUC). However, no cultivation-independent tests are available for the US_NmUC strains, and prior studies relied on microbial culture and genome sequencing to identify them. Here, we describe a PCR test that can identify the US_NmUC strains and distinguish them from commensal and invasive N. meningitidis strains as well as N. gonorrhoeae. Our SimpleProbe®-based real-time PCR assay targets a conserved nucleotide substitution in a horizontally acquired region of US_NmUC strain genomes. We applied the assay to 241 urine specimens whose microbial compositions had previously been determined by deep shotgun metagenomic sequencing. The assay detected the single US_NmUC positive case in this cohort, with no false positives. Overall, our simple and readily adaptable assay could facilitate investigation of the pathogenesis and epidemiology of the US_NmUC clade.

Sexually acquired syphilis: Historical aspects, microbiology, epidemiology, and clinical manifestations.

Syphilis is caused by infection with the spirochetal bacterium Treponema pallidum subsp. pallidum. It was first recognized in the late 15th century. Since 2000, the incidence of sexually acquired syphilis has increased substantially in the developed world, with men who have sex with men and persons living with HIV infection disproportionately affected. Clinical manifestations of syphilis are protean and often include mucocutaneous manifestations. The first article in this continuing medical education series reviews historical aspects, microbiology, epidemiology, and clinical manifestations of sexually acquired syphilis.

High willingness to use novel HIV and bacterial sexually transmitted infection partner notification, testing, and treatment strategies among gay and bisexual men.

OBJECTIVES: We sought to determine willingness of gay and bisexual men (GBM) to give HIV self-testing (HIVST) kits with patient-delivered partner therapy (PDPT) and engage in geosocial sexual networking (GSN) app-based partner notification. METHODS: A nationwide sample of GBM who self-tested HIV negative (n=786) were asked about their willingness to give recent sex partners (main and casual) PDPT with an HIVST kit (PDPT+HIVST) after hypothetical bacterial STI (BSTI) diagnosis. Men were also asked about their willingness to notify sexual partners met on GSN apps using an anonymous app function after BSTI diagnosis. We examined associations of relationship status and condomless anal sex with casual partners, recent BSTI diagnosis and perceived risk of HIV on PDPT+HIVST and anonymous app-based partner notification willingness (dichotomised) using binary logistic regressions, adjusting for age, race/ethnicity, education and US region. From the partner's perspective after receiving an app-based referral, frequency measures were used to report intentions for obtaining subsequent HIV/BSTI counselling and testing, engaging in HIVST if provided a free voucher, and obtaining BSTI treatment from a pharmacy with prescription voucher. RESULTS: Most (90.1%) were willing to give PDPT+HIVST to recent sex partners after STI diagnosis, and nearly all (96.4%) were willing to notify sex partners met online using an anonymous function within GSN apps. Regardless of casual partner condomless anal sex engagement, partnered GBM had higher odds of reporting willingness to give PDPT+HIVST compared with single men who recently engaged in condomless anal sex with a casual partner. If anonymously notified via an app, 92.5% reported they would likely obtain counselling and testing, 92.8% would engage in HIVST if provided a free voucher, and 93.4% would obtain treatment from a pharmacy with prescription voucher. CONCLUSIONS: GBM generally found novel partner notification, testing, and treatment strategies acceptable, indicating the need for feasibility and cost-effectiveness evaluations.

Assessment of rectal Chlamydia trachomatis viable load in women by viability-PCR.

OBJECTIVES: In recent years, studies have demonstrated frequent rectal Chlamydia trachomatis (CT) detection in women, irrespective of reported anal sex or rectal symptoms. However, the clinical relevance and public health implication of rectal CT detection in women remain under debate. Therefore, evaluating CT viability may provide more insight into the relevance of standard routine nucleic acid amplification test (NAAT)-positive results. METHODS: In this cross-sectional explorative study, a convenience sample of female patients at our STI clinic aged 18 years or older, diagnosed with vaginal and/or rectal CT, were invited to participate. On return for treatment, rectal CT-diagnosed women were instructed to self-collect rectal swab samples before being treated. Standard COBAS 4800 CT/NG routine NAAT testing was applied for CT diagnosis. Rectal viable CT load was evaluated by using viability-PCR (V-PCR). RESULTS: 53 women with rectal CT were included in this study; 86.8% (46/53) had a quantifiable rectal total CT load. Of women with quantifiable samples, 52.2% (24/46) had viable CT detected from rectal swabs by V-PCR, with a mean rectal viable CT load of 3.31 log10 CT/mL (range 1.16-6.22). No statistically significant difference (p=0.73) was observed in the mean rectal viable CT load of women with an indication for rectal testing (n=9) and without (n=15), 3.20 log10 CT/mL (range 2.06-4.36) and 3.38 log10 CT/mL (range 1.16-6.22), respectively. CT culture yielded positive test results from rectal swabs in 22.6% (12/53) of rectal CT NAAT-diagnosed women. Of women with viable rectal CT by V-PCR (n=24), 50% (12/24) were positive by CT culture. CONCLUSIONS: Overall, the detection of high rectal viable CT loads in this study indicates that rectal CT in some women might represent a currently ongoing infection rather than just the presence of remnant DNA from dead bacteria or only contamination from an active vaginal CT infection.

Manifestaciones coloproctológicas de las infecciones de transmisión sexual ocasionadas por Chlamydia Trachomatis, Neisseria Gonorrhoeae y Treponema Pallidum. Presentación casuística / Coloproctological manifestations of sexually transmitted infections caused by Chlamydia Trachomatis, Neisseria Gonorrhoeae and Treponema Pallidum. Case presentation

Introducción: Las infecciones transmisibles sexualmente (ITS) con afectación anorrectal constituyen un desafío pues las manifestaciones producidas por Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG) y Treponema pallidum (TP) son similares. Objetivo: Evaluar si las manifestaciones anorrectales debidas a CT, NG y TP asociadas al examen proctológico permiten diagnóstico certero, sin estudios complementarios. Pacientes y método: Estudio retrospectivo. Revisión de registros de pacientes atendidos en consultorio coloproctológico. Periodo: 01/08/2015-01/07/2016. Se incluyeron pacientes con diagnóstico de ITS anorrectal, excepto aquellos con HPV únicamente. A todos se les pesquisaron ITS mediante hisopado anal para CT por inmunofluorescencia y para estudio directo y cultivo de NG, VDRL para TP y además HIV. Variables: sexo, edad, HIV, sexo anal, uso de preservativo, motivo de consulta y resultado de estudios efectuados. Resultados: Treinta y cuatro pacientes (32 hombres). Edad mediana 31,5 años (rango: 19-65). Veinticinco pacientes HIV + (73,5%). Veintinueve pacientes (28 hombres) mantenían sexo anal. 91% no usaba preservativo adecuadamente. 65% tuvo una única infección (ITS pura). Se diagnosticaron 14 sífilis (8 puras), 14 clamidiasis (7 puras) y 11 gonococcias (7 puras). Co-infección entre ellas: 9% y con HPV: 26%. La úlcera fue la manifestación en 7/8 casos de sífilis puras (todas dolorosas, excepto una). El resto presentó síntomas variados (condilomas virales atípicos, secreción purulenta y proctorragia). Más del 50% de las gonococias puras (4/7) se manifestó con úlcera, sin embargo, el dolor estuvo presente siempre (8/8) y en tres se asoció secreción purulenta. En cambio, la mitad de los pacientes con clamidiasis puras, se manifestó con proctorragia causada por un tumor rectal/sigmoideo inflamatorio, clínicamente indistinguible de neoplasia maligna. Todos las sífilis y gonococias tuvieron correlato con las pruebas diagnósticas, no así las clamidiasis cuyo diagnóstico no pudo confirmarse en tres casos (37,5%), que respondieron al tratamiento empírico. Conclusión: NG y TP anorrectal provocaron mayormente síntomas similares a los de etiología no venérea y se requirió del laboratorio para el diagnóstico etiológico. La presencia de tumor con biopsia negativa para neoplasia maligna en pacientes de riesgo para ITS obliga a descartar clamidiasis. (AU)

Introduction: Sexually transmitted infections (STIs) are a challenge in medical consultation. The clinical manifestations of infection by Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG) and Treponema pallidum ( TP) share symptoms at anorectal level. This implies the need for a high index of suspicion for diagnosis, which is based on history, physical examination and laboratory tests that not always are accurate or available . Purpose: Assess whether clinical signs of anorectal infections by CT, NG and TP associated with proctologic exams, lead to an accurate etiologic diagnosis without the help of specific laboratory studies. Patients and methods: Observational, retrospective study, based on a review of records of patients treated at the outpatient clinic of the Hospital Fernandez (City of Buenos Aires) department of coloproctology, in the period between August 2015 and July 2016. Patients who underwent STI diagnosis were all considered, but to those whose only diagnosis was infection by human papilloma virus (HPV) were excluded from the analysis. All patients were tested after the three etiologies of STI (anal swab for CT study by immunofluorescence, swabbing for direct study, and cultivation of NG and TP VDRL) and HIV. Variables analyzed: sex, age, presence of HIV infection, practice of receptive anal sex, proper use of condoms, signs and symptoms that prompted the consultation, and results of diagnostic tests. Results: 34 patients (32 men) were included. Median age 31.5 years (range: 19-65, interquartile range: 26-37). Twenty-five patients (73.5%) were HIV+. Twenty-nine patients (28 men) remained receptive anal sex. 91% did not use condoms properly. 65% of infections were pure, without other STI asociada-. 14 cases of syphilis (8 pure), 14 Chlamydia (7 pure) and 11 gonococcias (7puras), including co-infection in 9% of cases, no evidence of a more frequent another co-infection diagnosed. Co-infection with HPV was detected in 9 (26%) cases. The ulcer was the sign in 7/8 cases of pure syphilis (all painful, except one). The rest is expressed by a variety of symptoms (atypical viral warts, purulent and bloody diarrhea). Similarly, just over 50% (4/7) of pure gonococcias demonstrated ulcer, but the pain was always present (8/8 of pure gonococcias) and three associated with purulent discharge. Instead of the ten patients with pure chlamydia, 50% manifested with bloody diarrhea caused by a rectal tumor / inflammatory sigmoid, clinically indistinguishable from malignancy. All cases of syphilis and gonococcal were correlated with diagnostic tests; not those whose diagnosis of chlamydial infection (confirmed in eight and was negative in three, 37.5%) who responded to empiric treatment indicated by the clinical suspicion. Conclusion: While this is a small series, it shows that the NG and TP in the anorectal location mostly caused symptoms similar to those of non-venereal ethology most of the times, and laboratory assistance for etiologic diagnosis was required. The presence of tumor with negative biopsy for malignancy in patients at risk for STIs, leads chlamydia to be ruled out. (AU)

Use of whole-genome sequencing to identify clusters of Shigella flexneri associated with sexual transmission in men who have sex with men in England: a validation study using linked behavioural data.

Since the 1970s, shigellosis has been reported as a sexually transmissible infection, and in recent years, genomic data have revealed the breadth of Shigella spp. transmission among global networks of men who have sex with men (MSM). In 2015, Public Health England (PHE) introduced routine whole-genome sequencing (WGS) of Shigella spp. to identify transmission clusters. However, limited behavioural information for the cases hampers interpretation. We investigated whether WGS can distinguish between clusters representing sexual transmission in MSM and clusters representing community (non-sexual) transmission to inform infection control. WGS data for Shigella flexneri from August 2015 to July 2017 were aggregated into single linkage clusters based on SNP typing using a range of SNP distances (the standard for Shigella surveillance at PHE is 10 SNPs). Clusters were classified as 'adult male', 'household', 'travel-associated' or 'community' using routine demographic data submitted alongside laboratory cultures. From August 2015 to March 2017, PHE contacted those with shigellosis as part of routine public-health follow-up and collected exposure data on a structured questionnaire, which for the first time included questions about sexual identity and behaviour. The questionnaire data were used to determine whether clusters classified as 'adult male' represented likely sexual transmission between men, thereby validating the use of the SNP clustering tool for informing appropriate public-health responses. Overall, 1006 S. flexneri cases were reported, of which 563 clustered with at least one other case (10-SNP threshold). Linked questionnaire data were available for 106 clustered cases, of which 84.0 % belonged to an 'adult male' cluster. At the 10-SNP threshold, 95.1 % [95 % confidence interval (CI) 88.0-98.1%] of MSM belonged to an 'adult male' cluster, while 73.2 % (95 % CI 49.1-87.5%) of non-MSM belonged to a 'community' or 'travel-associated' cluster. At the 25-SNP threshold, all MSM (95 % CI 96.0-100%) belonged to an 'adult male' cluster and 77.8 % (95 % CI 59.2-89.4%) of non-MSM belonged to a 'community' or 'travel-associated' cluster. Within one phylogenetic clade of S. flexneri, 9 clusters were identified (7 'adult male'; 2 'community') using a 10-SNP threshold, while a single 'adult male' cluster was identified using a 25-SNP threshold. Genotypic markers of azithromycin resistance were detected in 84.5 % (294/348) of 'adult male' cases and 20.9 % (9/43) of cases in other clusters (10-SNP threshold), the latter of which contained gay-identifying men who reported recent same-sex sexual contact. Our study suggests that SNP clustering can be used to identify Shigella clusters representing likely sexual transmission in MSM to inform infection control. Defining clusters requires a flexible approach in terms of genetic relatedness to ensure a clear understanding of underlying transmission networks.

A Systematic Literature Review of Reviews on the Effectiveness of Chlamydia Testing.

Chlamydia trachomatis is the most common bacterial sexually transmitted infection, causing significant morbidity and economic burden. Strategies like national screening programs or home-testing kits were introduced in some developed countries, yet their effectiveness remains controversial. In this systematic review, we examined reviews of chlamydia screening interventions to assess their effectiveness and the elements that contribute to their success to guide public policy and future research. We assessed English material published after 2000 in PubMed, the Cochrane Library, the British Nursing Index, Medical Database, and Sociological Abstracts, in addition to World Health Organization Global Health Sector Strategies, the European Center for Disease Prevention and Control guidelines, and the Prospective Register of Systematic Reviews. Systematic reviews that focused on chlamydia screening interventions were included. Using the socioecological model, we examined the levels of interventions that may affect the uptake of chlamydia screening. A total of 19 systematic reviews were included. Self-collection in home-testing kits significantly increased screening among girls and women 14-50 years of age. At the organizational level, using electronic health records and not creating additional costs facilitated testing. At the community level, outreach interventions in community and parent centers and homeless shelters achieved high screening rates. At the policy level, interventions with educational and advisory elements could result in significant improvements in screening rates.

Impact of specimen storage temperature and time on the implementation of GeneXpert® testing for sexually transmitted infections in resource-constraint settings.

Implementation of routine laboratory diagnostics is imperative to address the high burden of sexually transmitted infections (STI) in Sub-Saharan Africa. We demonstrate that logistical challenges of specimen storage, temperature and transport time are unlikely to impact on performance of routine STI diagnostics using the GeneXpert® platform implemented in these settings.