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Purpose. Bacterial vaginosis (BV) is a common clinical condition characterized by odorous vaginal discharge, vaginal itching and/or burning. BV can occur when vaginal lactobacilli are depleted and replaced by diverse anaerobic bacteria. We evaluated a commercial multiplex PCR (ATRiDA) for the diagnosis of BV. Methods. Cervicovaginal samples were included from women reporting urogenital symptoms and from women notified for sexually transmitted infections (STI) – who were not (necessarily) symptomatic. Clinical BV diagnoses were obtained from electronic patient files. The ATRiDA test measures the loads of Gardnerella vaginalis, Atopobium vaginae and Lactobacillus species in relation to overall bacterial load. The ATRiDA test outcome was compared to the clinical BV diagnosis and to vaginal microbiota composition, determined by 16SrRNA gene sequencing. Results. We included samples from 185 women reporting urogenital symptoms, of whom 81 had BV and 93 women who were notified for an STI, of whom 16 had BV. Overall, compared to the clinical BV diagnosis, the ATRiDA test demonstrated high sensitivity (96.9 %) and moderate specificity (70.2 %). The negative predictive value was high (>97.3). The positive predictive value differed by study group and was highest in women reporting urogenital symptoms (78.2 %). Sequencing showed that 54 % of women who had an ATRiDA BV-positive test outcome, but who were not clinically diagnosed with BV, had diverse anaerobic vaginal microbiota (asymptomatic vaginal dysbiosis). Conclusion. The ATRiDA test is a sensitive method for the detection of BV but, given the high occurrence of asymptomatic vaginal dysbiosis, a positive test outcome should be interpreted together with clinical symptoms.

IntroductionThis prospective study aimed to study the composition and structure of the vaginal microbiota prior to Chlamydia trachomatis infection.MethodsA nested case–control study was performed in 122 women, half of which acquired C. trachomatis within a year after their first visit. At the first visit, the composition and structure of vaginal microbial communities were analysed using 16S rRNA sequencing in the context of the sociodemographic and sexual risk behaviour information using logistic regression.ResultsFive vaginal community state types (CSTs) were identified. Four CSTs were dominated by Lactobacillus spp., of which L. crispatus (37%) and L. iners (33%) were the most common. One CST was characterised by the absence of Lactobacillus spp. (25%) and the presence of an array of strict and facultative anaerobes. Multivariate logistic regression analysis revealed that women with a L. iners-dominated CST had an increased risk of C. trachomatis infection (p=0.04; OR: 2.6, 95% CI 1.0 to 6.6).ConclusionsThe distribution of CSTs dominated by Lactobacillus spp. agreed with previous studies. However, the frequency of dysbiosis among Caucasian women was relatively high (24%). Having vaginal microbiota dominated by L. iners was associated with an increased risk for C. trachomatis infection. Therefore, we hypothesise that specific signatures of vaginal microbiota are indicative of increased host predisposition to acquiring STIs.

Background. Increasing evidence suggests that the cervicovaginal microbiota (CVM) plays an important role in acquiring sexually transmitted infections (STIs). Here we study the CVM in a population of women notified by a sex partner for Chlamydia trachomatis infection. Methods. We included 98 women who were contact-traced by C. trachomatis-positive sex partners at the STI outpatient clinic in Amsterdam, the Netherlands, and analyzed their cervicovaginal samples and clinical data. CVMs were characterized by sequencing the V3/V4 region of the 16S ribosomal RNA gene and by hierarchical clustering. Characteristics associating with C. trachomatis infection were examined using bivariable and multivariable logistic regression analysis. Results. The CVM was characterized for 93 women, of whom 52 tested C. trachomatis positive and 41 C. trachomatis negative. We identified 3 major CVM clusters. Clustered CVM predominantly comprised either diverse anaerobic bacteria (n = 39 [42%]), Lactobacillus iners (n = 32 [34%]), or Lactobacillus crispatus (n = 22 [24%]). In multivariable analysis, we found that CVM was significantly associated with C. trachomatis infection (odds ratio [OR], 4.2 [95% confidence interval {CI}, 1.2-15.4] for women with diverse anaerobic CVM and OR, 4.4 [95% CI, 1.3-15.6], for women with L. iners-dominated CVM, compared with women with L. crispatus-dominated CVM), as was younger age (OR, 3.1 [95% CI, 1.1-8.7] for those ≤21 years old) and reporting a steady sex partner (OR, 3.6 [95% CI, 1.4-9.4]). Conclusions. Women who tested positive for Chlamydia trachomatis infection after having been contact-traced by a chlamydiapositive partner were more likely to have CVM dominated by L. iners or by diverse anaerobic bacteria, than by L. crispatus.

Background Recognition of nosocomial outbreaks with antimicrobial resistant (AMR) pathogens and appropriate infection prevention measures are essential to limit the consequences of AMR pathogens to patients in hospitals. Because unrelated, but genetically similar AMR pathogens may circulate simultaneously, rapid high-resolution molecular typing methods are needed for outbreak management. We compared amplified fragment length polymorphism (AFLP) and whole genome sequencing (WGS) during a nosocomial outbreak of vancomycin-resistant Enterococcus faecium (VRE) that spanned 5 months. Methods Hierarchical clustering of AFLP profiles was performed using unweighted pair-grouping and similarity coefficients were calculated with Pearson correlation. For WGS-analysis, core single nucleotide polymorphisms (SNPs) were used to calculate the pairwise distance between isolates, construct a maximum likelihood phylogeny and establish a cut-off for relatedness of epidemiologically linked VRE isolates. SNP-variations in the vanB gene cluster were compared to increase the comparative resolution. Technical replicates of 2 isolates were sequenced to determine the number of core-SNPs derived from random sequencing errors. Results Of the 721 patients screened for VRE carriage, AFLP assigned isolates of 22 patients to the outbreak cluster. According to WGS, all 22 isolates belonged to ST117 but only 21 grouped in a tight phylogenetic cluster and carried vanB resistance gene clusters. Sequencing of technical replicates showed that 4–5 core-SNPs were derived by random sequencing errors. The cut-off for relatedness of epidemiologically linked VRE isolates was established at ≤7 core-SNPs. The discrepant isolate was separated from the index isolate by 61 core-SNPs and the vanB gene cluster was absent. In AFLP analysis this discrepant isolate was indistinguishable from the other outbreak isolates, forming a cluster with 92% similarity (cut-off for identical isolates ≥90%). The inclusion of the discrepant isolate in the outbreak resulted in the screening of 250 patients and quarantining of an entire ward. Conclusion AFLP was a rapid and affordable screening tool for characterising hospital VRE outbreaks. For in-depth understanding of the outbreak WGS was needed. Compared to AFLP, WGS provided higher resolution typing of VRE isolates with implications for outbreak management.

Introduction Of female visitors to the STI clinic in Amsterdam, 31% report use of vaginal cleansing products (including douches). Vaginal douching may negatively affect vaginal microbiota compositions (VM). We report the effect of intra-vaginal douching on the VM in a prospective study. Methods Through advertisements, we recruited 25 healthy women, aged 18–36 years, from 2015–2016. Participants were followed over 3 menstrual cycles and were instructed to use an intra-vaginal lactic-acid-containing douche 3 times a week during the 2nd cycle. Participants self-collected a median of 68 [IQR: 64–68] vaginal swabs. Baseline characteristics were collected through questionnaires. All participants kept a daily diary in which they reported douching, menstruation, sexual activity, etc. VM were assessed by 16S rRNA (V3-V4 region) sequencing. Associations between douching and VM were assessed by multivariable logistic regression, using generalised estimating equations to account for multiple observations within the same individual. Results As of December 2016, a median of 42 [IQR:40–44] vaginal swabs from 10/25 women were analysed. These 10 women had a median age of 25 years [IQR: 21.8–29.3], 9 women were Dutch-Caucasian, 9 used hormonal contraceptives and all were highly educated. At baseline, 8 women had lactobacilli-dominated VM ( Lactobacillus crispatus (n=6), L. iners (n=1) or L. iners / L. jensenii (n=1)) and 2 women had polybacterial Gardnerella vaginalis -containing VM (GV-VM). The latter 2 women continued to have GV-VM throughout the study period. The VMs of 2 women, dominated either by L. crispatus or L. iners at baseline, shifted to GV-VM during the 2nd cycle, which persisted in the 3rd cycle. Having GV-VM was more likely in the 2nd and 3rd cycle, compared to the 1 st cycle, after adjusting for sex and menses (odds ratio (OR)=1.7 (95% CI: 0.9–3.1) and OR=2.1 (95% CI: 0.7–6.1), respectively), though not statistically significanctly so. Conclusion Our interim analyses suggest that regular intra-vaginal douching may promote a shift from lactobacilli-dominated VM to GV-VM.

To evaluate hepatitis B virus (HBV) risk group vaccination in Amsterdam, which started in 1998, we examined 342 reported acute HBV-cases and sequenced 85 DNA isolates. The reported number of cases declined from 214 in 1992-1997 to 128 in 1998-2003, due to a decline in injecting drug users (IDU) and their heterosexual partners. Phylogenetic analyses showed that after 1998, the IDU cluster nearly disappeared, probably due to a decline in injecting. Acute HBV remained stable among men having sex with men; given their increased sexual risk behavior, vaccination has probably prevented an increase in their acute infections. Currently, 48-72% of the people who should be included in the program are still susceptible to HBV.

The effect of vaccination programs on transmission of infectious disease is usually assessed by monitoring programs that rely on notifications of symptomatic illness. For monitoring of infectious diseases with a high proportion of asymptomatic cases or a low reporting rate, molecular sequence data combined with modern coalescent-based techniques offer a complementary tool to assess transmission. Here, the authors investigate the added value of using viral sequence data to monitor a vaccination program that was started in 1998 and was targeted against hepatitis B virus in men who have sex with men in Amsterdam, the Netherlands. The incidence in this target group, as estimated from the notifications of acute infections with hepatitis B virus, was low; therefore, there was insufficient power to show a significant change in incidence. In contrast, the genetic diversity, as estimated from the viral sequence collected from the target group, revealed a marked decrease after vaccination was introduced. Taken together, the findings suggest that introduction of vaccination coincided with a change in the target group toward behavior with a higher risk of infection. The authors argue that molecular sequence data provide a powerful additional monitoring instrument, next to conventional case registration, for assessing the impact of vaccination.

Background Although published results are inconsistent, it has been suggested that tattooing and piercing are risk factors for HBV and HCV infections. To examine whether tattooing and piercing do indeed increase the risk of infection, we conducted a study among people with multiple tattoos and/or piercings in the Netherlands who acquired their tattoos and piercings in the Netherlands and/or abroad. Methods Tattoo artists, piercers, and people with multiple tattoos and/or piercings were recruited at tattoo conventions, shops (N = 182), and a biannual survey at our STI-outpatient clinic (N = 252) in Amsterdam. Participants were interviewed and tested for anti-HBc and anti-HCV. Determinants of HBV and HCV infections were analysed using logistic regression analysis. Results The median number of tattoos and piercings was 5 (IQR 2–10) and 2 (IQR 2–4), respectively. Almost 40% acquired their tattoo of piercing abroad. In total, 18/434 (4.2%, 95%CI: 2.64%–6.46%) participants were anti-HBc positive and 1 was anti-HCV positive (0.2%, 95%CI: 0.01%–1.29%). Being anti-HBc positive was independently associated with older age (OR 1.68, 95%CI: 1.03–2.75 per 10 years older) and being born in an HBV-endemic country (OR 7.39, 95%CI: 2.77–19.7). Tattoo- and/or piercing-related variables, like having a tattoo or piercing in an HBV endemic country, surface percentage tattooed, number of tattoos and piercings etc., were not associated with either HBV or HCV. Conclusions We found no evidence for an increased HBV/HCV seroprevalence among persons with multiple tattoos and/or piercings, which might be due to the introduction of hygiene guidelines for tattoo and piercing shops in combination with the low observed prevalence of HBV/HCV in the general population. Tattoos and/or piercings, therefore, should not be considered risk factors for HBV/HCV in the Dutch population. These findings imply the importance of implementation of hygiene guidelines in other countries.