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Hypothesis/aims of study: Infection of the amniotic cavity and placenta is one of the leading causes of adverse pregnancy outcomes. In the majority of cases, intra-amniotic infection is associated with the normal microbiota of the lower urogenital tract. The aim of the study was to explore the relationships between the placental inflammatory changes, vaginal microbiota and labor course. Study design, materials and methods: We examined 124 women at 37-41 weeks of gestation. The vaginal discharge at admission was taken for microbiological evaluation, with the delivered placenta sent for histological examination. Results: In 17.7% of cases, histological examination of the placenta revealed inflammatory changes. A statistically significant correlation was noted between the placental inflammatory changes and Staphylocossus spp. presence in the vaginal discharge at admission (р = 0.0004). The placental inflammatory changes were associated with the membrane rupture to delivery interval more than 6 hours (р = 0.01) and the labor duration more than 7 hours (р = 0.0004). Prelabor rupture of membranes did not significantly affect the placental inflammatory changes (p = 1.0). Conclusion: Predisposing factors for the development of ascending bacterial infection of the placenta are an abnormal vaginal microbiota with the presence of opportunistic bacteria before delivery, a long membrane rupture to delivery interval, and a prolonged labor.

Evaluation of circulating group B streptococcus (GBS) strains is important to assess the potential effects of GBS prevention strategies in a certain region. This study aimed at estimating the distributions of capsular polysaccharide (CPS) types and pilus profiles, and the rates of antimicrobial resistance among GBS strains isolated from colonized pregnant women and newborns in 2010-2011 and 2017-2018 in St. Petersburg, Russia. A total of 261 GBS isolates have been investigated. Antibiotic susceptibility testing was performed using the disc-diffusion method. CPS types and pilus profiles were determined by using PCR. Over the 9-year period, the resistance of GBS to both erythromycin and clindamycin has significantly increased, exceeding 30% in 2017-2018. The most prevalent CPS types were Ia, III, and V. Significant shifts were observed in the frequency of CPS types III (decreased) and V (increased), which resulted in a significant reduction (from 77 to 63%) in the potential coverage by a trivalent vaccine (including serotypes Ia, Ib, and III), whereas that of a pentavalent vaccine (including serotypes Ia, Ib, II, III, and V) remained largely unchanged (approximately 95%). The most common pilus profiles were PI-1/2a, PI-2a, and PI-1a/2b, and pilus genotype distribution has not changed with time. High and steadily growing resistance of perinatal GBS strains to clindamycin requires restricting its use to penicillin-allergic women at high risk for anaphylaxis and testing the GBS strains for their susceptibility to this antibiotic. A pentavalent CPS-based vaccine covers the vast majority of perinatal GBS strains in Russia.

The large majority of studies investigating associations between bacterial vaginosis (BV) and sexually transmitted infections (STIs) have been conducted among predominantly young women with high risk for STIs. Since a risky sexual behavior is a significant risk factor for both STIs and BV, this creates a bias toward an increased association between BV and STIs. This study evaluated associations between BV-associated vaginal microbiota and STIs (Chlamydia trachomatis, Mycoplasma genitalium, Trichomonas vaginalis, and Neisseria gonorrhoeae) in a population of women with low risk for STIs and investigated STI outcomes depending on the dominating Lactobacillus species. Repository cervicovaginal samples collected from reproductive-age women from January 2014 to February 2019 were characterized for vaginal microbiota types and the STIs using multiplex real-time PCR assays. In total, 95 STI-positive and 91 STI-negative samples were included. A significant, age-independent association between BV-associated vaginal microbiota and the presence of C. trachomatis, M. genitalium, and T. vaginalis infections was identified (age-adjusted odds ratios 2.92 [95% confidence interval (CI) 1.24–7.03], 2.88 [95% CI 1.19–7.16], and 9.75 × 107 [95% CI 13.03-∞], respectively). Normal vaginal microbiota dominated by Lactobacillus crispatus, L. gasseri, or L. jensenii was a strong protective factor against C. trachomatis and/or M. genitalium infections, whereas L. iners-dominated microbiota was not significantly associated with C. trachomatis and/or M. genitalium positivity. The results of the present study confirm that STI prevention strategies should include interventions that also reduce the incidence of BV and promote a protective vaginal microbiota in both high- and low-risk women.

Hypothesis/aims of study. Vulvovaginal candidiasis (VVC) is one of the most common inflammatory diseases of the vagina. Herewith, a synergistic effect of lactobacilli and antifungal drugs in VVC treatment is suggested. In this regard, it seems relevant to study the effect of antimycotics used to treat VVC on the enhanced L. casei rhamnosus 35 (LCR35) probiotic strain. This study was aimed to determine the sensitivity to antimycotic drugs of the probiotic LCR35 strain included in the Lactoginal® vaginal capsules, as well as the sensibility of the genus Candida yeast under separate and joint cultivation with lactobacilli. Study design, materials and methods. LCR35 was cultured on a MRS agar. Identification of bacteria grown on the medium, as well as clinical isolates of C. albicans and C. parapsilosis , isolated from the vagina, was performed by MALDI-TOF mass spectrometry. The sensitivity of yeast was determined on a modified Muller-Hinton agar with 40% glucose and methylene blue. Discs with fluconazole, voriconazole, itraconazole, ketoconazole, clotrimazole, and nystatin were used. Interpretation of growth inhibition zones was carried out according to EUCAST-2018 or according to the recommendations of the disc manufacturer. Results. The probiotic LCR35 strain is resistant to antimycotics that are effective against C. albicans and C. parapsilosis . When determining the sensibility of C. albicans and C. parapsilosis to antimycotics in separate and joint cultivation with LCR35, no differences were found in the degree of yeast-like fungi growth suppression. Conclusion. The probiotic LCR35 strain, which is part of the Lactoginal® vaginal capsules, is resistant to antimycotic drugs and does not affect the sensitivity of the tested yeast-like fungi to antimycotics.

Background Infections with human papillomavirus (HPV) types 16 and 18 account for ~70% of invasive cervical cancers but the degree of protection from naturally acquired anti‐HPV antibodies is uncertain. We examined the risk of HPV infections as defined by HPV DNA detection and cervical abnormalities among women >25 years in the Human Papilloma VIrus Vaccine Immunogenicity ANd Efficacy trial's (VIVIANE, NCT00294047) control arm. Methods Serum anti‐HPV‐16/18 antibodies were determined at baseline and every 12 months in baseline DNA‐negative women (N = 2687 for HPV‐16 and 2705 for HPV‐18) by enzyme‐linked immunosorbent assay (ELISA) from blood samples. HPV infections were identified by polymerase chain reaction (PCR) every 6‐months, and cervical abnormalities were confirmed by cytology every 12 months. Data were collected over a 7‐year period. The association between the risk of type‐specific infection and cervical abnormalities and serostatus was assessed using Cox proportional hazard models. Results Risk of newly detected HPV‐16‐associated 6‐month persistent infections (PI) (hazard ratio [HR] = 0.56 [95%CI:0.32; 0.99]) and atypical squamous cells of undetermined significance (ASC‐US+) (HR = 0.28 [0.12; 0.67]) were significantly lower in baseline seropositive vs baseline seronegative women. HPV‐16‐associated incident infections (HR = 0.81 [0.56; 1.16]) and 12‐month PI (HR = 0.53 [0.24; 1.16]) showed the same trend. A similar trend of lower risk was observed in HPV‐18‐seropositive vs ‐seronegative women (HR = 0.95 [0.59; 1.51] for IIs, HR = 0.43 [0.16; 1.13] for 6‐month PIs, HR = 0.31 [0.07; 1.36] for 12‐month PIs, and HR = 0.61 [0.23; 1.61] for ASC‐US+). Conclusions Naturally acquired anti‐HPV‐16 antibodies were associated with a decreased risk of subsequent infection and cervical abnormalities in women >25 years. This possible protection was lower than that previously reported in 15‐ to 25‐year‐old women.

Background Bacterial vaginosis (BV) is a vaginal disorder characterized by a depletion of the normal lactobacillus-dominant microbiota and overgrowth of mainly anaerobic bacteria. Objectives The study aimed to evaluate the distribution and abundance of the Gardnerella vaginalis clades and sialidase A gene in vaginal samples from Russian women, and investigate if the G. vaginalis sialidase A gene count detects an abnormal vaginal microbiota characteristic of BV more accurately than G. vaginalis load. Methods Vaginal samples from 299 non-pregnant patients of gynecological clinics were examined using Nugent scores and G. vaginalis clade and sialidase A gene quantitative real-time polymerase chain reactions (PCRs). Discriminatory power for BV microbiota was evaluated with receiver operating characteristic (ROC) analysis. Results The vaginal microbiota was characterized by Nugent scores as normal, intermediate, and BV microbiota in 162, 58, and 79 women, respectively. G. vaginalis clades 1, 2, 3, 4, and the sialidase A gene were detected in 56% (51–62%), 40% (34–45%), 20% (16–25%), 94% (91–96%), and 70% (64–75%) of vaginal samples, respectively. The frequency and abundance of clades 1, 2, 4, and the sialidase A gene as well as clade multiplicity were significantly associated with abnormal microbiota. The sialidase A gene was present in all multi-clade samples, in all single-clade samples comprising clades 1, 2, and 3, and in four of 84 (5% [2–12%]) samples comprising clade 4 only. Total G. vaginalis load showed significantly higher discriminatory power for abnormal microbiota than sialidase A gene count (areas under ROC curves 0.933 vs. 0.881; p = 0.0306). Conclusions Quantifying all four G. vaginalis clades discriminates between BV microbiota and normal microbiota more accurately than measuring G. vaginalis sialidase A gene. Clade 4 is strongly associated with BV microbiota, despite most clade 4 strains lacking the sialidase A gene.

ObjectivesAntimicrobial resistance in Mycoplasma genitalium (MG) is a poorly surveyed and controlled global health concern. We evaluated the first commercial dual resistance assay, AmpliSens M. genitalium-ML/FQ-Resist-FL assay, for detection of potential macrolide and quinolone resistance-associated mutations (MRAMs and QRAMs, respectively) and estimated the prevalence of these mutations in MG in St. Petersburg, Russia.MethodsUrogenital samples positive (n=145 from 2007 to 2020) and negative (n=56 from 2021) for MG in routine diagnostics were retrospectively analysed using the AmpliSens M. genitalium-ML/FQ-Resist-FL assay (Central Research Institute of Epidemiology, Moscow, Russia) and Sanger sequencing for validation.ResultsThe AmpliSens M. genitalium-ML/FQ-Resist-FL assay detected potential MRAMs and QRAMs with sensitivities of 100% (CI95% 83.9 to 100) and 92.3% (CI95% 66.7 to 99.6) and specificities of 99.2% (CI95% 95.6 to 100) and 100% (CI95% 97.2 to 100), respectively, in clinical specimens with ≥1000 MG geq/mL. In total, MRAMs were detected in 13.8% (CI95% 9.1 to 20.3) of samples, with 23S rRNA A2058G being the most prevalent mutation (45.0% (CI95% 25.8 to 65.8)). QRAMs were found in 9.0% (CI95% 5.3 to 14.7) of samples, with S83I the most frequent mutation (53.8% (CI95% 29.1 to 76.8)). Dual resistance was observed in 5.5% (CI95% 2.8 to 10.5) of samples. Potential MRAM and dual resistance rates significantly increased over time: from 0% in 2007–2008 to 25% (ptrend=0.0009) and 10% (ptrend=0.0447), respectively, in 2018–2020. QRAM rate appeared to increase (from 0% to 13%), but significance was not reached (ptrend=0.0605).ConclusionsThe rapid increase in MG antimicrobial resistance in St. Petersburg, especially prominent for MRAMs, necessitates implementation of macrolide resistance-guided therapy in Russia. The first commercial dual resistance assay, AmpliSens M. genitalium-ML/FQ-Resist-FL assay, was sensitive and specific for detection of potential MRAMs and QRAMs and could be valuable in macrolide resistance-guided therapies and possibly for surveillance of QRAMs. International surveillance of antimicrobial resistance-associated mutations in MG, further research into clinical relevance of several parC mutations and novel treatments are essential.

The objective of our study was to explore the possible graphene impact on microorganism growth as well as on laboratory animal overall condition.Graphene did not contain any microorganisms, does not destroy erythrocytes placed within the artificial nutritional medium, graphene lattice did not add any properties to the quartz glasses which could allow the Proteus spread all over its surface. It was also established that graphene did not show any native antibacterial impact. No significant reaction was noticed in animals after graphene administration to laboratory rats neither at the injection spot nor at the overall level.Our data confirm the applicability of graphene both in scientific and practical biomedical purposes.

Background: Urinary tract infections (UTIs) are common in women, and during pregnancy can cause significant morbidity. Growing and greatly varying antimicrobial resistance (AMR) of Enterobacteriaceae, responsible for most UTIs, necessitates regular local AMR surveillance. In obstetric population, where beta-lactams are the mainstay for treatment of severe UTIs, particular focus should be placed on beta-lactam resistance. This study aimed to evaluate AMR rates and frequency of extended spectrum beta-lactamase (ESBL) and carbapenemase genes in uropathogenic Enterobacteriaceae among reproductive-age women in St. Petersburg, Russia.Materials/methods: Urine samples were collected from consecutive reproductive-age women, who attended the D.O. Ott Research Institute of Obstetrics, Gynecology and Reproductology from October 2017 to November 2019, and cultured according to routine procedures. Susceptibility to antibiotics and ESBL production was determined using the disc diffusion method according to the European Committee on Antimicrobial Susceptibility Testing (EUCAST) guidelines. All urine samples and Enterobacteriaceae isolates were tested for ESBL and carbapenemase genes using real-time PCR.Results: Enterobacteriaceae were detected in 91 (56 pregnant and 35 non-pregnant) of 119 (76%) included women. The vast majority of Enterobacteriaceae strains were susceptible to nitrofurantoin, fosfomycin and meropenem (99-100%). The frequency of strains susceptible to penicillins and cephalosporins ranged from 59% to 82%; 78% of strains were susceptible to ciprofloxacin. ESBL production was phenotypically detected in 15 (16%) Enterobacteriaceae strains, with CTX-M genes revealed in all cases. In all corresponding urine samples, CTX-M genes were also detected. The remaining 104 urine samples were negative for CTX-M genes. In none of the isolates and urine samples, carbapenemase genes were present.Conclusions: The frequency of ESBL producing Enterobacteriaceae was relatively high (16%), with CTX-M genes detected in all cases in both urine and urine cultures. Rapid PCR detection of CTX-M genes directly in urine samples from women with pyelonephritis can be valuable for timely informing treatment choices.

Hypothesis/aims of study. A lack of timely diagnosis and treatment of inflammatory diseases of the lower genital tract in a woman can lead to serious reproductive disorders. The detection of inflammation is an important component of the diagnosis of these diseases. This study was undertaken to compare methods for detection of local inflammation in women with complaints of vaginal discharge. Study design, materials, and methods. A total of 87 women having vaginal discharge were enrolled in the study. For the detection of inflammation, vaginal discharge was examined using microscopy and reverse transcriptase polymerase chain reaction (the ImmunoQuantex® test, DNA-Technology Ltd., Moscow, Russia). Results. Vaginal infections caused by opportunistic microorganisms were detected in 27 women (31%), as follows: candidal vulvovaginitis (CVV) in 10 patients (11.5%), aerobic vaginitis (AV) in 14 patients (16.1%), and bacterial vaginosis (BV) in 14 patients (16.1%), with 8 (9.2%) of them having BV combined with CVV, AV or both infections. The results of the two methods fully agreed in the analysis of samples from women with CVV and AV as vaginal infections characterized by local inflammation. Testing samples from women with BV, which is a non-inflammatory syndrome, as well as women without vaginal infections resulted in significant discrepancies between the two methods. The sensitivity and specificity of the ImmunoQuantex® test compared to the microscopic method were 85.7% and 48.9%, respectively. Conclusion. The new reverse transcriptase polymerase chain reaction based test for the evaluation of expression profiles of selected genes of immune response enables to assess the local immune status in the vagina. Determination of the presence or absence of inflammatory reaction in the vagina can be used as a diagnostic criterion for resolving discrepancies between laboratory data and clinical manifestations, as well as for choosing effective therapy.

Hypothesis/aims of study. The present analysis was undertaken to summarize current knowledge about molecular properties of group B streptococci (GBS), emphasizing potential targets of vaccines against neonatal GBS infection. Study design, materials, and methods. This review is based on articles published mainly in the last ten years. Results. Epidemiological data on serotypes, multilocus sequence types, clonal complexes of GBS and their relationship are presented. Genetic events in GBS populations indicate significant obstacles to vaccine development. We described key properties of major GBS virulence factors, such as capsular polysaccharide, pili, and cell adhesion molecules, as well as results of experimental immunization on their basis. Conclusion. The population of invasive GBS strains is molecularly and genetically heterogeneous, which complicates selection of vaccine targets. Capsular switching, a low level of immunogenicity and variability of population composition are the most important factors that necessitate the accumulation and monitoring of molecular epidemiological data.

Hypothesis/aims of study. Dysbiotic disorders of the vagina are one of the main risk factors of miscarriage. The analysis of vaginal microbiota during pregnancy in women having a history of miscarriage may contribute to understanding the importance of specific types of vaginal flora in the development of processes resulting in spontaneous abortion or preterm birth. This study was undertaken to evaluate the vaginal flora in the 1st trimester of pregnancy in women having a history of miscarriage. Study design, materials, and methods. To participate in the study, 160 individuals in the 1st trimester of pregnancy with and without a history of miscarriage were invited. Group 1 included 100 women with a history of miscarriage, group 2 consisted of 60 women without a history of miscarriage. The vaginal flora was analyzed with microscopic, bacteriological, and quantitative real-time PCR methods. Results. In patients with a history of miscarriage, the frequency of dysbiotic disorders of the vaginal flora was three times higher (11.0% and 3.3%) and pH value of the vaginal fluid was significantly higher. Along with a significant reduction of the proportion of the Lactobacillus flora in the 1st trimester of pregnancy in patients having a history of miscarriage, there was a significant increase in the abundance of opportunistic microorganisms, mostly aerobic flora, such as Enterobacteriaceae, Streptococcus spp., and Staphylococcus spp. Furthermore, there was a significant predominance of Ureaplasma spp. and Candida spp. in women with a miscarriage history. Conclusion. The vaginal microbiota in the 1st trimester of pregnancy in women having a history of miscarriage is characterized by higher pH values of the vaginal fluid, the prevalence of opportunistic microorganisms, a decrease in the abundance of the Lactobacillus flora, and a higher concentration of aerobic bacteria, Ureaplasma spp., and Candida spp. These findings indicate dysbiotic disorders of the vaginal microbiocenosis in women having a history of miscarriage, which can lead to late miscarriage and pretem birth.

Background. Pelvic inflammatory disease (PID), including chronic endometritis (CE), is a factor leading to endometrial receptivity disorder and in vitro fertilization (IVF) failures. The role of microorganisms in colonizing the endometrium, the pathogenesis of chronic inflammation, and associated infertility require further study. Aims: To determine the taxonomic profile of the endometrial microbiota in different grades of CE in patients with a history of IVF failure. Materials and methods. This study included 107 patients with ≥ 1 IVF failures. Depending on the severity of CE, and according to the morphological analysis of endometrial biopsy, the patients were categorized into three groups: group I (n = 14), patients without signs of CE; group II (n= 20); patients with signs of mild CE; and group III (n = 73),patients with signs of moderate and severe CE. All the patients underwent Pipelle biopsy of the endometrium according to our modified technique on days 18–23 of the menstrual cycle. Species and quantitative composition of the endometrial microbiota were investigated using real-time polymerase chain reaction (PCR). Results. Compared to groups II and III, Lactobacillus spp. (50% and 46.6% versus 85.7%) and Eubacterium spp. (25% and 35.6% versus 78.6%) were detected significantly more frequently in the endometrial microflora of group I patients, respectively. Simultaneously, significantly more bacteria of the families Enterobacteriaceae (50% and 64.4% versus 35.7%, respectively) and Streptococcus spp. (55% and 56.2% versus 7.1%, respectively) were found in the patients of groups II and III compared to group I. The abundance of lactobacilli and Eubacterium spp. have also been shown to be significantly higher in the endometrial samples of women in group I than in those of the women in groups II and III, whereas the abundance of enterobacteria and streptococci are significantly higher in women with CE than in women without CE. Conclusions. In patients without signs of CE, Lactobacillus spp. and Eubacterium spp. prevail in the endometrial microbiota. Chronic inflammation in the endometrium is associated with an increase in the frequency and abundance of Streptococcus spp., Staphylococcus spp., and bacteria of the family Enterobacteriaceae.

Background. Bacteriuria caused by group B streptococci (GBS) is a major risk factor for neonatal GBS associated pathology and a risk factor for pregnancy complications such as preterm birth and preterm rupture of membranes. Objective. Clinical and microbiological rationale for the prophylaxis of obstetrical and perinatal pathology associated with group B streptococci. Methods. A total of 496 pregnant women who made their first prenatal visit within first 12 weeks of gestation were invited to participate. As clinical samples, mid-stream urine, vaginal swabs, rectal swabs from women and inguinal swabs, urine, meconium from newborn infants were used. GBS detection was performed using culture and PCR. Results. The frequency of GBS detection was 16.3%. GBS associated bacteriuria was detected in 9.8% of the women. The women having GBS in urine at the beginning of pregnancy were significantly more often colonized with the microorganism late in pregnancy in comparison with the women with GBS-colonization of the vagina and/or rectum. Microbiological efficiency of antibiotic therapy of GBS associated bacteriuria using amoxicillin/clavulanic acid exceeded 70%. Clinical efficiency of the therapy consisted in the reduction of preterm birth and preterm rupture of membranes by 25% and 40%, respectively. Conclusions. Antibiotic therapy of GBS associated bacteriuria results in a decrease of pregnancy complication, which necessitate its use. GBS-colonization of the vagina and rectum in pregnant women is transitory therefore antenatal screening for GBS aimed to prevent neonatal GBS-infection should be performed late in pregnancy or before delivery.

Background. Bacterial vaginosis (BV) is disturbance of the vaginal microbiota, characterized by displacement of lactobacilli with anaerobic bacteria and capable of adversely affecting women’s reproductive health. In the development of BV, a wide spectrum of bacteria substantially differing in their properties is involved. Grouping vaginal bacterial communities into clusters, or types of microbiocenosis, might contribute to understanding of pathogenic mechanisms and elaboration of effective tools for diagnostics and therapy of the disease. Aim. Determination and comparative analysis of clusters of vaginal bacterial communities in norm and in BV. Materials and methods. Women of reproductive age were enrolled in the study. For the diagnosis of BV, the Nugent score was used. Vaginal swab samples from all women were analyzed with the test Femoflor-16, intended for evaluation of the vaginal microbiocenosis using multiplex quantitative real-time PCR. Two-step cluster analysis was applied for grouping bacterial communities. Differences between the clusters were evaluated using pairwise comparisons. Results. Of 280 women enrolled in the study, 172 had normal microflora, 27 – intermediate microflora, 81 – BV. In cluster analysis, 270 samples valid in PCR testing were included. All the vaginal bacterial communities were grouped into 4 clusters. Cluster 1 (n = 171) included cases when the vaginal microflora consisted mostly of lactobacilli. Cluster 2 (n = 11) encompassed cases of domination of aerobic microflora: Enterobacteriaceae, Streptococcus and Staphylococcus. Clusters 3 (n = 57) and 4 (n = 31) were connected with BV and included cases of prevailing of facultative anaerobes (Gardnerella vaginalis, Atopobium vaginae) and obligate anaerobes (Sneathia/Leptotrichia/Fusobacterium, Megasphaera/Veillonella/Dialister, Lachnobacterium/Clostridium), respectively. Nearly all cases of cluster 1 belonged to the category of normal microflora of the Nugent score. The majority of bacterial communities of cluster 2 matched intermediate microflora, cluster 3 – BV category with a score of 7 or 8, cluster 4 – BV category with a score of 9 or 10. The clusters differed significantly in vaginal рН, with the highest values observed for cluster 4. Conclusions. Vaginal bacterial communities are grouped into 4 main clusters, characterized by domination of lactobacilli, aerobes, facultative anaerobes or obligate anaerobes. The clusters belong to different categories of the Nugent score and differ significantly in vaginal pH.

Background. Group B streptococci (GBS) are opportunistic bacteria capable of causing severe infections in pregnant women and newborn infants. For effective prevention of obstetrical and perinatal GBS-associated pathology, investigation into epidemiology of GBS carriage and GBS-associated diseases, as well as elaboration of accurate and rapid methods for GBS detection are necessary. Objectives. Validation of a molecular test for the detection of GBS and evaluation of frequency and risk factors of colonization of pregnant women with this microorganism. Methods. For validation of PCR test for the detection of GBS DNA (AmpliSens Streptococcus agalactiae-screen-titer-FL, Central Research Institute of Epidemiology, Moscow), 1496 clinical samples from 650 women and 112 newborn infants submitted for routine GBS culture were used. For evaluation of frequency and risk factors of GBS-colonization during pregnancy, clinical samples (urine, vaginal and rectal swab samples) from 496 women at their first prenatal visit before 12 weeks of gestation were used. GBS testing was performed using culture and the validated PCR test. For evaluation of risk factors, binomial logistic regression was used. Results. PCR method for GBS DNA detection showed high analytical sensitivity (3 ∙ 102 copies/ml) and specificity (no cross-reactions with other microorganisms). Diagnostic sensitivity and specificity of the test (81 and 97.6%, respectively) were comparable with those of culture (77.6 and 100%, respectively). GBS was detected in 16.3% of pregnant women. Testing of rectal samples in addition to vaginal samples enabled to detect 66.7% more cases of GBS carriage. Women aged 18 to 22 years had a higher risk of GBS colonization during pregnancy than older women (23 to 32 years). At the same time, women who had their sexual debut at an older age (after 18) were significantly more often colonized during pregnancy than women started their sex life at a younger age (before 18). Conclusion. PCR method for GBS detection has high sensitivity and specificity and can be used as an alternative to culture. Simultaneous testing of vaginal and rectal samples increases the sensitivity of revealing colonized women by two thirds in comparison with testing vaginal samples only. Younger age and later sexual debut are independent risk factors for GBS colonization during pregnancy.

Introduction. The study is in vitro experiment, aimed to determine antibiotic susceptibility of microorganisms isolated from the urogenital tract of women to the combination of substances included in the preparation Terzhinan. Methods. In total, 516 microorganism strains isolated from the vagina of reproductive age women have been tested: Candida spp. (n = 83), bacteria of the family Enterobacteriaceae (n = 138), Streptococcus agalactiae (n = 72), Enterococcus spp. (n = 147), Staphylococcus spp. (n = 37), Actinomyces urogenitalis (n = 7), Gardnerella vaginalis (n = 27), Atopobium vaginae (n = 5). Suspensions of each strain in a volume of 0.5 ml (0.5 according to McFarland) were applied onto culture medium (Muller-Hinton Agar or 5% Blood Muller-Hinton Agar). Furthermore, serial dilutions of Terzhinan were applied with a calibrated loop onto three sectors. Petri dishes were then incubated at 37 °С for 24 h. The results were evaluated visually, by measuring growth inhibition zones. Results. The majority of Candida albicans isolates were susceptible (S) to the preparation, using both undiluted and diluted (10 and 100 times) preparation. Terzhinan without dilution and in the dilution 1:10 was 100% effective against all tested bacteria of the family Enterobacteriaceae. Susceptibility of gram positive bacteria to the antibiotics included in Terzhinan, was 74.1%. All isolates of staphylococci, including Staphylococcus aureus, were susceptible. The frequency of susceptible strains of Streptococcus agalactiae and Enterococcus spp. was 70%. All clinical isolates of Gardnerella vaginalis and Atopobium vaginae were susceptible to the combinations of antibiotics included in the preparation Terzhinan, which is in agreement with data on its high efficiency in the treatment of bacterial vaginosis. Conclusions. In in vitro experiment, the combined preparation Terzhinan showed high activity against major groups of microorganisms isolated from the vagina including pathological conditions, such as bacterial vaginosis and vulvovaginitis of different etiology.

Background. Bacterial vaginosis is disturbance of the balance of the vaginal microflora, associated with a number of infectious diseases of the urogenital tract and adverse pregnancy outcomes. In this country, for the detection of vaginal dysbiotic conditions, the test Femoflor-16 (DNA-Technology, Moscow) is widely used, however interpretation algorithms of this test do not include the category of BV. Aim. The study aimed to elaborate diagnostic criteria for the detection of BV using Femoflor-16 test. Materials and methods. Women of reproductive age addressing a gynecologist with vaginal discharge were enrolled in the study. For clinical diagnosis of BV, the Amsel criteria were used, laboratory analysis for BV was performed via microscopic investigation of vaginal discharge using the Nugent score. Samples of vaginal discharge from all women were analyzed with the test Femoflor-16, intended for characterizing vaginal microbiocenosis using multiplex quantitative real-time PCR. Results. A total of 280 women were included in the study. BV was diagnosed in 86 women (31%) using the Amsel criteria, and in 81 women (29%) using the Nugent score. All groups of anaerobic bacteria included in Femoflor-16 test were shown to be associated with BV, with the exception of bacteria of the genus Mobiluncus, which are detected together with phylogenetically related but not BV-associated bacteria of the genus Corynebacterium. A low amount of lactobacilli (< 10% of total bacterial load) coupled with an elevated amount of Gardnerella vaginalis/Prevotella bivia/Porphyromonas (> 1%) and/or Eubacterium (> 2%) and/or Sneathia/Leptotrichia/Fusobacterium (> 0.1%) and/or Megasphaera/Veillonella/Dialister (> 0.1%) and/or Lachnobacterium/Clostridium (> 0.1%) and/or Peptostreptococcus (> 0.1%) and/or Atopobium vaginae (> 0.2%) detected BV with a sensitivity of 99% and specificity of 93%. Conclusions. Criteria for BV diagnosis using the test Femoflor-16 have been elaborated, which enable to detect BV or exclude it with a sensitivity of 99% and specificity of 93%. These criteria for BV and criteria of the test manufacturers for severe anaerobic dysbiosis determine to a large extent the same category of the vaginal microbiocenosis.

Introduction. In healthy women of reproductive age, the vaginal microflora is represented mainly by lactobacilli. They provide a barrier function, preventing the propagation of opportunistic pathogens and the colonization of the vagina by pathogenic microorganisms. It is shown that the lactobacillary microflora of the vagina is very diverse, but not all species of lactobacilli can provide reliable protection of the female organism. Objective: to characterize the species and quantitative composition of vaginal lactobacilli of women of reproductive age in norm and in dysbiosis. Material and methods. The study involved 123 patients from polyclinic departments of the D.O. Ott Reasearch Institute of Obstetrics, Gynecology and Reproductology. For analysis of clinical material (vaginal discharge) for lactobacilli and other microorganisms, quantitative real-time PCR was used. Results. The most common types of lactobacilli are Lactobacillus jensenii, L. Iners, L. crispatus, L. vaginalis and L. gasseri. Species diversity of lactobacilli (detection of ≥2 species) is observed much more often in women with physiological microbiocenosis than in women with vaginal dysbiosis. Our study confirms that L. crispatus is the dominant species of the vaginal biotope of healthy women, while in dysbiosis the species L. gasseri and L. Iners are most often identified.

The present paper describes for the first time a case of using the preparation of vaginal lactobacilli Lactoginal (strain Lactobacillus rhamnosus Lcr35) for correction of vaginal microflora disturbance, associated with cervical cerclage in pregnant woman with a history of pregnancy miscarriage. Administration of the probiotic lactobacilli preparation up to the delivery resulted in steady normalization of the vaginal miсrobiocenosis, which might have, to a certain extent, contributed to its normal course and delivery. The obtained results are discussed in the context of literature data.

Summary Background Although the risk of human papillomavirus (HPV) infection is greatest in young women, women older than 25 years remain at risk. We present data from the VIVIANE study of the HPV 16/18 AS04-adjuvanted vaccine in adult women after 7 years of follow-up. Methods In this phase 3, double-blind, randomised controlled trial, healthy women older than 25 years were enrolled (age stratified: 26–35 years, 36–45 years, and ≥46 years). Up to 15% in each age stratum had a history of HPV infection or disease. Women were randomly assigned (1:1) to receive HPV 16/18 vaccine or aluminium hydroxide control, with an internet-based system. The primary endpoint was vaccine efficacy against 6-month persistent infection or cervical intraepithelial neoplasia grade 1 or greater (CIN1+) associated with HPV 16/18. We did analyses in the according-to-protocol cohort for efficacy and total vaccinated cohort. Data for the combined primary endpoint in the according-to-protocol cohort for efficacy were considered significant when the lower limit of the 96·2% CI around the point estimate was greater than 30%. For all other endpoints and cohorts, data were considered significant when the lower limit of the 96·2% CI was greater than 0%. This study is registered with ClinicalTrials.gov, number NCT00294047. Findings The first participant was enrolled on Feb 16, 2006, and the last study visit took place on Jan 29, 2014. 4407 women were in the according-to-protocol cohort for efficacy (n=2209 vaccine, n=2198 control) and 5747 women in the total vaccinated cohort (n=2877 vaccine, n=2870 control). At month 84, in women seronegative for the corresponding HPV type in the according-to-protocol cohort for efficacy, vaccine efficacy against 6-month persistent infection or CIN1+ associated with HPV 16/18 was significant in all age groups combined (90·5%, 96·2% CI 78·6–96·5). Vaccine efficacy against HPV 16/18-related cytological abnormalities (atypical squamous cells of undetermined significance and low-grade squamous intraepithelial lesion) and CIN1+ was also significant. We also noted significant cross-protective efficacy against 6-month persistent infection with HPV 31 (65·8%, 96·2% CI 24·9–85·8) and HPV 45 (70·7%, 96·2% CI 34·2–88·4). In the total vaccinated cohort, vaccine efficacy against CIN1+ irrespective of HPV was significant (22·9%, 96·2% CI 4·8–37·7). Serious adverse events related to vaccination occurred in five (0·2%) of 2877 women in the vaccine group and eight (0·3%) of 2870 women in the control group. Interpretation In women older than 25 years, the HPV 16/18 vaccine continues to protect against infections, cytological abnormalities, and lesions associated with HPV 16/18 and CIN1+ irrespective of HPV type, and infection with non-vaccine types HPV 31 and HPV 45 over 7 years of follow-up. Funding GlaxoSmithKline Biologicals SA.

A local inflammatoMicrobial invasion of amniotic cavity and subsequent infection process are the leading cause of adverse pregnancy outcomes. Intra-amniotic infection can be caused by both pathogenic and opportunistic microorga­nisms and viruses, but in the vast majority of cases intra-amniotic infection is associated with the normal microflora of different body sites – the vagina, intestines, mouth and respiratory tract. In the paper, current views about the role of infection factor in the adverse pregnancy outcomes, sources and mechanisms of intra-amniotic infection are summarized.

The article describes the history of the Laboratory of Microbiology of the D.O. Ott Research Institute of Obstetrics, Gynecology since its foundation till present days. The paper summarizes information on the studies of outstanding Russian obstetricians-gynecologists conducted at the end of the 19th to the beginning of the 20th centuries, using innovative for those times bacteriological methods. At present, the Laboratory of Microbiology is a high-technology department of the Institute, equipped with advanced instruments and staffed with highly proficient personnel. The staff of the Laboratory of Microbiology carefully keeps the history and adheres to the traditions of research work initiated by distinguished scientists in the past.

Background. Trichomoniasis is accompanied by substantial colonization of the genital tract by various opportunistic microorganisms and reduced amount of lactobacilli. Both processes have a negative impact on the reproductive health and fetal development. Today, however, in the treatment of trichomoniasis insufficient attention is paid to the restoration of the normal vaginal microflora. Aim. An attempt was made to assess the vaginal microbiocenosis of patients with trichomoniasis using traditional and novel molecular methods. Matherials and Methods. Archived vaginal samples from 37 women with trichomoniasis were used, which were placed on slides, fixed in the flame and stained according the Gram method and by methylene blue and stored in boxes within last 15 years. The preparations were evaluated using microscopy and the nucleic acid amplification based test Femoflor and tests for the detection of DNA of sexually transmitted agents. Results. According to the microscopic examination, disorders of the vaginal microbiocenosis were detected in 89.2% of patients with trichomoniasis. C. trachomatis was detected in 8.1% of the samples. Obligate anaerobic microorganisms were determined in all samples, accounting for 8% to 100% of the total bacterialthe majority of the women. Facultative anaerobic microflora was also found in all the patients. Lactobacilli were detected in 8.1% of the women, and mycoplasma – in 37.8%. Conclusions. It was shown that in women with trichomoniasis there are often disturbances of the normal vaginal flora, manifested as the presence of a variety of opportunistic bacteria, accompanied by an inflammatory response. Almost half of the preparations showed bacterial vaginosis. Noteworthy, there was high prevalence of Mycoplasma hominis. For the first time, a technique of DNA testing in the materials placed on glass slides after prolonged storage was applied.

Microbial biofilms are a problem in many areas of medicine. When the vaginal ecological system is disturbed, well-structured polymicrobial biofilm, covering the vaginal epithelium and consisting of anaerobic bacteria, is formed, which may lead to the development of recurrent bacterial vaginosis. During an ascending infection and the development of chronic endometritis, bacterial biofilms are detected in the endometrium. Biofilms formed by bacteria residing in the urogenital tract of woman, may result in dysfunctions of the reproductive system, in-vitro fertilization failures, complications of pregnancy, adverse pregnancy outcomes. In the paper, literature data regarding causes of of biofilm formation, methods of detection and treatment of biofilm infections are reviewed.

Introduction. Abnormal vaginal microflora in the first trimester of pregnancy is a risk factor for miscarriage. Opportunistic microorganisms predominate among microorganisms causing microflora disturbance. Aim. The aim of the study was to evaluate the vaginal microflora in the first trimesterф Introduction. Abnormal vaginal microflora in the first trimester of pregnancy is a risk factor for miscarriage. Opportunistic microorganisms predominate among microorganisms causing microflora disturbance. Aim. The aim of the study was to evaluate the vaginal microflora in the first trimester of pregnancy in women having history of pregnancy loss. Matherials and Methods . Real-time polymerase chain reaction, microscopic and cultural methods were used to examine 60 pregnant women having history of pregnancy loss and 10 pregnant women with uncomplicated obstetric history. Results. Disorders of the vaginal microflora were observed only in women having history of pregnancy loss. The lactoflora in both groups was shown to consist most frequently of 3 Lactobacillus species: Lactobacillus crispatus, L. jensenii, and L. iners. However, the species L. iners were detected only in the group of women with history of pregnancy loss. Non-Lactobacillus microbiota was significantly more often observed in pregnant women having history of pregnancy loss. Facultative and obligate anaerobes were detected in both groups. The concentration of Ureaplasma spp. and Mycoplasma hominis in women in both groups had no significant differences. Conclusions. The results of this study indicate that abnormal microbiota is more often observed in women having history of pregnancy loss. of pregnancy in women having history of pregnancy loss. Matherials and Methods. Real-time polymerase chain reaction, microscopic and cultural methods were used to examine 60 pregnant women having history of pregnancy loss and 10 pregnant women with uncomplicated obstetric history. Results. Disorders of the vaginal microflora were observed only in women having history of pregnancy loss. The lactoflora in both groups was shown to consist most frequently of 3 Lactobacillus species: Lactobacillus crispatus, L. jensenii, and L. iners. However, the species L. iners were detected only in the group of women with history of pregnancy loss. Non-Lactobacillus microbiota was significantly more often observed in pregnant women having history of pregnancy loss. Facultative and obligate anaerobes were detected in both groups. The concentration of Ureaplasma spp. and Mycoplasma hominis in women in both groups had no significant differences. Conclusions. The results of this study indicate that abnormal microbiota is more often observed in women having history of pregnancy loss.

Introduction. Bacterial vaginosis (BV) is associated with a number of reproductive health disorders, therefore timely and accurate diagnosis of this condition is exceedingly important. Objective. Comparison of effectiveness of clinical and laboratory diagnostics of BV in women with vaginal discharge. Material and methods. In total, 318 patients addressing gynecological clinics with complaints about vaginal discharge participated in the study. Clinical diagnostics of BV was performed in the clinics participating in patient enrollment in accordance with their clinical practice. For laboratory diagnostics, microscopy of Gram stained smears according to the Nugent method and quantitative real-time PCR were used. Sensitivity and specificity of clinical diagnostics of BV and the molecular method were evaluated using the Nugent method as reference standard. Results. With the Nugent method, BV was diagnosed in 27% of women, with real-time PCR — in 37% of women. Using clinical signs of BV, the condition was diagnosed in 91% women. Sensitivity and specificity of the real-time PCR were 97% and 87%, respectively. Sensitivity of clinical diagnostics was 100%, but specificity was only 17%. Conclusions. Diagnostics of BV based only on the presence of vaginal discharge leads to false positive results and requires laboratory confirmation. The molecular method has a high sensitivity and satisfactory specificity for BV diagnosis and can be used as an alternative to the Nugent method.

Introduction. Urinary tract infections are the most common infections in obstetrics and gynecology. Bacteriological method to investigate urine is laborious and time-consuming, therefore development of accurate and rapid methods for the detection of significant bacteriuria is important. Objective. Evaluation of quantitative real-time PCR based approach for the detection of significant bacteriuria in pregnant women. Material and methods. A retrospective investigation of mid-stream urine samples obtained from pregnant women was performed. Urine culture was performed using quantitative method, and a case was considered as significant bacteriuria if ≥ 105 CFU/ml were detected. Urine samples were analyzed for main uropathogens / groups of uropathogens using quantitative multiplex real-time PCR. Diagnostic characteristics of PCR were computed relative to the results of urine culture. Results. In total, 896 urine samples were tested. Of them, significant bacteriuria was found in 28 cases (3%). The frequency of detection of Escherichia coli was 50%, Enterococcus spp. — 25%, Klebsiella spp. — 7%, Proteus spp. and S. saprophyticus 4% each, Streptococcus spp. — 14%. Sensitivity and specificity of the detection of significant bacteriuria using quantitative real-time PCR for the majority of bacterial species / groups were 99% to 100%. Sensitivity and specificity of the quantitative real-time PCR based method were 96% and 98%, respectively. Conclusions . Prevalence of significant bacteriuria among pregnant women is 3%. Half of the uropathogens isolated from pregnant women with bacteriuria are E. coli. Sensitivity and specificity of quantitative PCR for the detection of significant bacteriuria are 96% and 98%, respectively.

Introduction. Bacterial vaginosis (BV) is the primary cause of pathological vaginal discharge in women of reproductive age. Gardnerella vaginalis and Atopobium vaginae are considered key components of the vaginal microflora in BV. Etiology, pathogenesis and modes of transmission of BV are actively studied, however these questions still remain unanswered. Objective: investigate predictor factors of BV in women with vaginal discharge. Material and methods. In total, 318 women were included. As clinical material, vaginal samples were used. BV was diagnosed using the Nugent method. For quantitative determination of G. vaginalis and A. vaginae DNA, real-time PCR was used. Behavioral and anamnestic data were obtained from questionnaire filled out by the patients. Results. BV was diagnosed in 27 % of women. G. vaginalis and A. vaginae DNA was detected, respectively, in 93 % and 83 % of patients with BV, 73 % and 59 % - with intermediate microflora, 52 % and 38 % - with normal microflora. Difference between the three types of microflora in the frequency and concentrations of these microorganisms were statistically significant. Detection of G. vaginalis and A. vaginae were significant predictor factors of BV (OR 12.2; 95 % CI 5.1-29.4 and OR 7.9; 95 % CI 4.2-14.9, respectively), with chances to diagnose BV being manifold increased when clinically significant concentrations of these bacteria were detected (≥3×106 and ≥8×105 DNA copies/ml for G. vaginalis and A. vaginae, respectively). Detection of clue cells in Gram stained preparations was shown to be the strongest BV predictor (OR 765.6; 95 % CI 99.6-5883.2). Conclusions. BV is diagnosed in more than one fourth of women with vaginal discharge. Detection of G. vaginalis and A. vaginae, especially in clinically significant concentrations, and clue cells in Gram stained preparations are significant predictor factors of BV.

The article presents the results of a survey of 84 women diagnosed with bacterial vaginosis on the basis of criteria of Amsel. The frequency of detection of various microorganisms/groups of microorganisms present in the vaginal microbiocenosis when different diagnostic criteria for bacterial vaginosis Amsel. Structure and diversity vaginal microbiocenosis assessed using molecular PCR in real time. Bacterial vaginosis is polyetiology syndrome. However, the search for individual bacteria and their combinations in bacterial vaginosis is an actual problem of modern researchers.

Urinary tract infections (UTIs) are among the most common infectious diseases in women, and are the most frequent cause of infectious complications of pregnancy. This paper reviews current scientific and methodical literature on UTIs in obstetrics and gynecology. Aspects of clinical importance of UTIs (epidemiology, clinical manifestations, complications), their etiology, antimicrobial resistance of UTIs agents were discussed, and current recommendations on diagnostics and treatment of UTIs were summarized. Special attention was paid to UTIs in pregnancy.

Enterococcus are opportunistic bacteria and are members of the normal microflora of the gastrointestinal tract of humans and animals. The prevalence of enterococcal infections in pregnant women is approximately 28 %. Enterococcus, colonizing the urogenital tract pregnant women mostly do not show pathogenic effect, however, it is known that they can lead to the development of such complications of pregnancy as threatened preterm labor, placental insufficiency, hypotrophy of the fetus. Of complications in childbirth most common in premature labor, untimely discharge of amniotic fluid, fetal hypoxia. According to the literature cause of neonatal bacteremia and sepsis in 10 % of cases are enterococcus. Thus, for the prevention of complications of pregnancy, delivery and perinatal outcomes in the allocation of Enterococcus urogenital tract to undergo antibacterial therapy, given the sensitivity to antibiotics.

Background: Elevated antibody titers to Chlamydia trachomatis are detected in up to 70% of women with tubal factor infertility (TFI). In order to evaluate the influence of past chlamydial infection on the results of IVF treatment, a prospective cohort study was performed.Materials and methods: A total of 242 women with TFI were enrolled in the study. Serum samples (n = 242) and follicular fluid (n = 197) were analyzed using enzyme-linked immunosorbent assay for anti-C. trachomatis IgG, IgA, IgM and anti-chlamydial heat shock protein 60 kDa IgG.Results: “Poor response” to ovarian stimulation was two times more frequent in seropositive subjects (22.9% versus 13.3%, p = 0.039). Seropositive and seronegative women had similar IVF outcomes, expressed by clinical pregnancy (20.5% versus 25.0%), life birth (13.1% versus 20.0%) and miscarriage rates (36.0% versus 16.7%, p = 0.092). Incidence of missed abortion after IVF was higher in women with previous exposure to C. trachomatis (28.0%) as compared to th...

Chlamydia trachomatis is the world's most prevalent bacterial sexually transmitted infection and leading infectious cause of blindness, yet it is one of the least understood human pathogens, in part due to the difficulties of in vitro culturing and the lack of available tools for genetic manipulation. Genome sequencing has reinvigorated this field, shedding light on the contemporary history of this pathogen. Here, we analyze 563 full genomes, 455 of which are novel, to show that the history of the species comprises two phases, and conclude that the currently circulating lineages are the result of evolution in different genomic ecotypes. Temporal analysis indicates these lineages have recently expanded in the space of thousands of years, rather than the millions of years as previously thought, a finding that dramatically changes our understanding of this pathogen's history. Finally, at a time when almost every pathogen is becoming increasingly resistant to antimicrobials, we show that there is no evidence of circulating genomic resistance in C. trachomatis., Funding agencies:Sanger Institute through the Wellcome Trust 098051 Australian National Health and Medical Research Council 1060768

Group B streptococcal infections (GBS) are the leading cause of sepsis, pneumonia and meningitis in infants during the first three months of life. GBS infections presenting within the first week of life are designated early-onset diseases, and those affecting infants between one week and three months - late-onset diseases. Intrapartum intravenous administration of antibiotics to women at high risk of transmitting the microorganism to the infants is currently considered the most effective method of prevention of early-onset GBS infections, and universal screening for GBS in late pregnancy - as the most effective method to detect women to whom antibiotic prophylaxis is indicated. Vaccination of pregnant women appears to be a promising approach to prevent both early- and late-onset GBS infections in infants, as well as GBS-associated diseases in women.

The study investigates colonization of the genitourinary tract and rectum in pregnant women by Streptococcus agalactiae in early pregnancy, as well as pregnancy outcomes for mother and infant depending on the use of antibiotic prophylaxis. The frequency of GBS colonization was 15.9 %, with the detection rate in urine being 8.6 %, in the vagina - 3.5 %, in the rectum - 10 %. The efficiency of amoxicillin/clavulanic acid administered in the II trimester was 66.7 %. Pregnancy outcomes in women receiving and those not receiving antibiotic prophylaxis were studied. In all women receiving amoxicillin/clavulanic acid term labor occurred, with healthy full-term infants born. In one woman not receiving antibiotic prophylaxis there was preterm labor at 34 weeks of gestation.

Bacterial vaginosis (BV) is the most common vaginal disorder in women of reproductive age. This review presents literature data regarding a role of BV in adverse reproductive health outcomes, describes current methods used for BV diagnosis and issues of treatment of this disease in pregnant women.

Urinary tract infections (UTIs) represent the most common bacterial infections and often complicate pregnancy. UTIs in pregnancy are classified by site of bacterial proliferation as follows: asymptomatic bacteriuria, cystitis, pyelonephritis. Screening for asymptomatic bacteriuria is a standard of obstetrical care and is included in most international guidelines. The urine microbiologic culture is considered the gold standard for laboratory diagnosis of UTIs. Molecular methods enable to rapidly detect, identify and quantitate causative agents of UTIs in urine and are effective alternative to traditional bacteriological methods.

Background and objective Resistance in the sexually transmitted bacterium Mycoplasma genitalium to all recommended therapeutic antimicrobials have rapidly emerged. However, to date, internationally reported resistance surveillance data for M. genitalium strains circulating in Eastern Europe are entirely lacking. The aim of this study was to estimate the prevalence of macrolide and fluoroquinolone resistance-associated mutations in M. genitalium in four cities in Russia and one in Estonia, 2013–2016. Materials and methods Consecutive urogenital samples found positive for M. genitalium during diagnostic testing were retrospectively analyzed for resistance-associated mutations in the 23S rRNA and parC genes using pyrosequencing and conventional Sanger sequencing, respectively. Results In total, 867 M. genitalium positive samples from 2013–2016 were analyzed. Macrolide resistance-associated mutations were detected in 4.6% of the samples from Russia (0.7–6.8% in different cities) and in 10% of the samples from Estonia. The mutations A2059G and A2058G were highly predominating in both Russia and Estonia, accounting together for 90.9% of the cases positive for nucleotide substitutions in the 23S rRNA gene. The rates of possible fluoroquinolone resistance-associated mutations were 6.2% in Russia (2.5–7.6% in different cities) and 5% in Estonia. The mutations S83I and S83N were the most frequent ones in Russia (24.4% each), whereas D87N highly predominated in Estonia (83.3% of all fluoroquinolone resistance-associated mutations). Approximately 1% of the samples in both countries harbored both macrolide and possible fluoroquinolone resistance-associated mutations, with A2058G and S83I being the most frequent combination (37.5%). Conclusions The prevalence of macrolide and fluoroquinolone resistance-associated mutations in M. genitalium was 4.6% and 6.2%, respectively, in Russia, and 10% and 5%, respectively, in Estonia. Despite the relatively low rates of macrolide and fluoroquinolone resistance in these countries, antimicrobial resistance surveillance and testing for resistance-associated mutations in M. genitalium positive cases would be valuable.

Chlamydia trachomatis is the world's most prevalent bacterial sexually transmitted infection and leading infectious cause of blindness, yet it is one of the least understood human pathogens, in part due to the difficulties of in vitro culturing and the lack of available tools for genetic manipulation. Genome sequencing has reinvigorated this field, shedding light on the contemporary history of this pathogen. Here, we analyze 563 full genomes, 455 of which are novel, to show that the history of the species comprises two phases, and conclude that the currently circulating lineages are the result of evolution in different genomic ecotypes. Temporal analysis indicates these lineages have recently expanded in the space of thousands of years, rather than the millions of years as previously thought, a finding that dramatically changes our understanding of this pathogen's history. Finally, at a time when almost every pathogen is becoming increasingly resistant to antimicrobials, we show that there is no evidence of circulating genomic resistance in C. trachomatis.

The laboratory diagnosis of sexually transmitted infections in many Eastern European countries remains suboptimal. The main objective of the present evidence-based guidelines is to provide comprehensive information regarding the laboratory diagnosis of infections caused by Trichomonas vaginalis in East European countries. In particular, the present guidelines recommend: (i) to encourage examination of the wet mounts of vaginal exudates, instead of stained smears, at all clinical settings; (ii) nucleic acid amplification tests (NAATs) or culture could be employed if no trichomonads are detected on microscopic examination of the wet preparation and there is a strong indication of infection and (iii) the use of NAATs is encouraged in screening, using non-invasive specimens, or high volume testing situations. In the absence of internationally recognized commercial NAAT systems, tests developed in-house should be validated using obtainable international standards and quality assured strictly. Individual East European countries may be required to make minor national adjustments to these guidelines as a result of lack of accessibility to some reagents or equipment, or laws in a specific country.

Background Diagnosis of Mycoplasma genitalium is entirely based on nucleic acid amplification tests (NAATs). In Russia, several M. genitalium polymerase chain reaction (PCR) assays have been developed; however, any evaluation of their performance has never been performed. Objective To assess the performance of five PCRs developed and currently used in Russia for diagnosis of M. genitalium. Materials and methods Vaginal swabs and first voided urine samples (FVUs) from 281 females and urethral swabs and FVUs from 125 males were analysed using three conventional PCRs and two real-time PCRs developed by three Russian companies. As reference tests, a real-time PCR targeting the MgPa adhesin gene was used; positive results were confirmed by two conventional PCRs targeting the 16S rRNA gene and MgPa gene, respectively. For evaluation of detection limits and analytical specificities, a blinded control panel consisting of dilutions of six strains of M. genitalium and 14 other Mycoplasma species was tested. Results The prevalence of M. genitalium was 2.5% among females and 9.6% among males. The highest sensitivity (71.4-100% in different specimens) was exhibited by one real-time PCRs. Conventional PCRs from two manufacturers failed to detect M. genitalium in any of the seven positive female FVUs. All tests had a 100% clinical specificity; however, one cross-reacted with Mycoplasma pneumoniae. Conclusions Only one of the five Russian PCRs displayed reasonable sensitivity for all specimen types, but the specificities of all assays were high. Accordingly, improvements regarding sensitivity of all the tests are needed. However, larger studies, including other populations, evaluating these assays are crucial.

The present guidelines aim to provide comprehensive information regarding the laboratory diagnosis of infections caused by Chlamydia trachomatis in East European countries. These recommendations contain important information for laboratory staff working with sexually transmitted infections (STIs) and/or STI-related issues. Individual East European countries may be required to make minor national adjustments to these guidelines as a result of lack of accessibility to some reagents or equipment, or laws in a specific country.

BACKGROUND: The numbers and performance characteristics of laboratories providing sexually transmitted infection (STI) diagnostic services, as well as the rates of morbidity due to STIs in St. Petersburg, Russia, remain largely unknown. OBJECTIVE: The aim of the present study was to evaluate the range, quality and availability of diagnostic services for several non-viral STIs (Chlamydia trachomatis, Neisseria gonorrhoeae, Treponema pallidum and Trichomonas vaginalis) in St. Petersburg during the period September 2005 to June 2006. METHODS: Survey data focusing on organization and performance characteristics of STI diagnostic services were assessed using questionnaires, telephone interviews and site visits. RESULTS: A total of 118 laboratories providing STI diagnostic services were identified. Of the surveyed laboratories, 54% (64 of 118) diagnosed syphilis, 81% (96 of 118) gonorrhoea, 80% (94 of 118) trichomoniasis and 49% (58 of 118) chlamydial infections. Although most of the laboratories could provide a presumptive diagnosis for syphilis, most of the N. gonorrhoeae and T. vaginalis testing of women did not adhere to international recommendations. Of the laboratories with the capacity to diagnose C. trachomatis infection, 69% still used serological testing (enzyme-linked immunosorbent assay) to detect antibodies to C. trachomatis. CONCLUSIONS: Overall, the diagnostic methods used to establish a laboratory diagnosis, the system of case reporting, the training of laboratory personnel and the level of interlaboratory communication clearly require improvement. This study represents the first step in a process of evaluation of the laboratory support for STI services and the establishment of an interlaboratory network in St. Petersburg.

This study aimed to assess the laboratory diagnosis of Neisseria gonorrhoeae in St. Petersburg, Russia. In total, 334 consecutive symptomatic patients were enrolled. Cervical and urethral specimens from women (n=286) and urethral specimens from men (n=48) were analyzed by microscopy, culture and two in-house NAATs, i.e. polymerase chain reaction (PCR) and nucleic acid sequence-based amplification (NASBA), developed in Russia. All N. gonorrhoeae-positive samples were confirmed using porA pseudogene and 16S rRNA gene sequencing. All methods displayed 100% specificity, i.e. positive predictive values of 100%. Compared to the PCR (most sensitive method in the present study), in women the sensitivity of both microscopy and culture was 31.8%, and that of NASBA was 90.9%. In men, microscopy, culture and NASBA displayed a sensitivity of 75%, 50% and 100%, respectively. The negative predictive values of microscopy, culture, and NASBA were 97.3%, 97.3%, and 99.6% in women, and 97.8%, 95.7%, and 100% in men, respectively. According to the PCR, the prevalences of N. gonorrhoeae were 4.5% (women) and 8.3% (men). In conclusion, both the investigated Russian NAATs displayed a high sensitivity and specificity. However, in general the diagnosis of gonorrhoea in Russia is suboptimal and crucially requires validation, improvements and quality assurance.

Purpose Little is known about the susceptibility of posterior segment tissues, particularly the human retinal pigment epithelium (hRPE), to Chlamydia trachomatis. The purpose of the study was to investigate the possibility of infecting the hRPE with Chlamydia trachomatis, and to examine the infectivity of different Chlamydia trachomatis clinical isolates for hRPE cells and the hRPE cell response to the infection. Methods Cultured hRPE and McCoy cells were inoculated with eight Chlamydia trachomatis (serovar E) clinical isolates at multiplicity of infection (MOI) of 2.0 or 0.3. To detect Chlamydia trachomatis, samples were stained immunohistochemically with anti-major outer membrane protein antibodies at 24h, 48h, and 72h postinoculation (PI). The changes in the expression of signaling molecules and proteins of cytoskeleton and extracellular matrix in hRPE cells were examined immunohistochemically. Results All eight clinical isolates demonstrated ability to infect hRPE cells. At equal MOI of 0.3, the infectivity of Chlamydia trachomatis clinical isolates for RPE culture was found to be at least as high as that for McCoy cell culture. At 24h PI, the percentage of inclusion-containing cells varied from 1.5 ± 0.52 to 14.6 ± 3.3% in hRPE cell culture infected at MOI of 2.0 against 0.37 ± 0.34 to 8.9 ± 0.2% in McCoy cell culture infected at MOI of 0.3. Collagen type I, collagen type IV, basic fibroblast growth factor, transforming growth factor-beta and interleukin–8 expression at 48h PI were maximally increased, by 2.1-, 1.3-, 1.5-, 1.5- and 1.6-fold, respectively, in the Chlamydia trachomatis-infected compared with control hRPE cell culture specimens (P < 0.05). Conclusions This study, for the first time, proved the possibility of infecting hRPE cultured cells with Chlamydia trachomatis, which leads to proproliferative and proinflammatory changes in the expression of signaling molecules and extracellular matrix components.

In the present study, the performance of the cell culture method, two non-Russian direct immunofluorescence (DIF) assays, and three different in-house polymerase chain reaction (PCR) tests used in St. Petersburg, Russia, for detection of Chlamydia trachomatis in urogenital specimens was evaluated. A total of 650 patients were examined and it was most disquieting that previous C. trachomatis positivity with Russian DIF assays could - 7 days later - be confirmed only in 26% of the women and 30% of the men. Overall, the highest diagnostic sensitivity was obtained using PCR analysis. However, the sensitivity varied significantly: from 79% to 100% between the different PCR assays, sex of the patients, and type of samples. The highest sensitivity was obtained for female vaginal and male urine samples (100%). The specificity of the PCR assays varied from 97% to 100%. The sensitivity of cell culture and both the examined DIF assays was low, i.e. it varied from 46% to 56% and 55% to 75%, respectively. Meanwhile, cell culture was 100% specific and the DIFs showed a specificity varying from 99% to 100%. In conclusion, in a Russian perspective, adequate in-house PCR methods may be used quite effectively for detection of C. trachomatis in invasive as well as non-invasive clinical material. Simultaneous analysis of two different specimens from women resulted in a significantly increased detection rate of C. trachomatis. Nevertheless, in Russia the need for optimization and quality assurance of diagnostic methods for C. trachomatis, especially Russian DIF assays, has to be emphasized.