Evaluation of polymerase chain reaction assays for the diagnosis of Trichomonas vaginalis infection in Russia.

BACKGROUND: In Russia, the microscopy- and culture-based diagnostics of trichomoniasis is mainly suboptimal. Recent years, domestically produced diagnostic PCR assays have been implemented; however, any evaluation of these PCRs has never been internationally reported. OBJECTIVE: To assess the performance characteristics of PCR assays developed and currently used in Russia to detect Trichomonas vaginalis. MATERIALS AND METHODS: Five PCR assays were assessed on 448 samples (317 vaginal and 131 male urethral) collected from symptomatic attendees of youth centres (n = 415) and patients of a dermatovenereological dispensary that were previously diagnosed with trichomoniasis (n = 33). As reference assay, a sensitive and specific real-time multiplex PCR was used. RESULTS: T. vaginalis DNA was detected in five (all females) of the 415 patients of youth centres (1.2%). All 33 patients previously diagnosed at the venereological dispensary proved to be true positive. For 445 (99.3%) of these 448 samples identical results were obtained by all PCRs, 35 positive and 410 negative. The three discordant samples were positive in all PCRs except one conventional PCR assay. The sensitivities of the PCRs were 94.3-100% and 66.7-100% for vaginal and urethral swabs, respectively. All evaluated assays were 100% specific. The detection limits of the different PCRs ranged from 0.1 to 5 genome equivalents per reaction. CONCLUSION: The PCR assays currently used in Russia for the detection of T. vaginalis have in general high sensitivities and excellent specificities for both vaginal samples and urethral samples from males.

Sexual behaviours, knowledge and attitudes regarding safe sex, and prevalence of non-viral sexually transmitted infections among attendees of youth clinics in St. Petersburg, Russia.

BACKGROUND: Adolescents and young adults are at increased risk of sexually transmitted infections (STIs). Knowledge of STI prevalence and risk factors are essential tools to elaborate preventive strategies. However, internationally reported studies on epidemiology of STIs among the youth in Russia are mainly lacking. OBJECTIVES: To ascertain sexual behaviours, knowledge and attitudes about safe sex and prevalence and correlates with STIs in attendees of youth clinics in St. Petersburg, Russia. METHODS: A total of 301 women and 131 men, who self-referred for STI testing, completed a questionnaire and were screened for Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium, and Trichomonas vaginalis using nucleic acid amplification tests. RESULTS: The overall STI prevalence was 16.9%, and similar in the female patients and male patients (15.6% and 19.8% respectively). C. trachomatis, N. gonorrhoeae, M. genitalium and T. vaginalis were detected in 13%, 2.5%, 4.6% and 1.2% of the attendees respectively. The men displayed riskier sexual behaviours and worse knowledge and attitudes regarding safe sex compared to the women, with the most distinguishing features being younger age at first intercourse (P < 0.0005), higher numbers of sex partners during lifetime (P = 0.001) and latest 6 months (P < 0.0005), more frequently consuming alcohol (P < 0.0005), poorer knowledge of STI/HIV prevention measures (P < 0.0005), and less positive attitudes towards safe sex (P = 0.001). However, no significant predictors of STI positivity were found in the men. In the women, the strongest predictors of STI positivity were young age (15-19 years) and multiple sex partners (≥ 2) during latest 6 months. CONCLUSIONS: The overall prevalence of STIs among users of STI services at youth clinics in St. Petersburg was high. Comprehensive epidemiological data on STI prevalence and sexual behaviour correlates are necessary to initiate new and strengthen existing STI prevention programmes for the youth, in Russia as well as in many other settings.

Recommended antimicrobial treatment of uncomplicated gonorrhoea in 2009 in 11 East European countries: implementation of a Neisseria gonorrhoeae antimicrobial susceptibility programme in this region is crucial.

BACKGROUND: Antimicrobial resistance (AMR) in Neisseria gonorrhoeae (gonococcus) is a major problem worldwide. Quality-assured and quality-controlled AMR surveillance data on gonococci globally are crucial for public health purposes. In East European countries, knowledge regarding gonococcal AMR and its prevalence is limited. OBJECTIVES: To ascertain the recommendations for antimicrobial treatment of uncomplicated gonorrhoea in 11 East European countries, valuable information for introducing an international gonococcal AMR surveillance programme. METHODS: A questionnaire was used to collect information regarding the types, doses and manufacturers of the antimicrobials recommended for gonorrhoea treatment in all countries. RESULTS: Ceftriaxone (250-1000 mg, intramuscularly (IM)×1) was reported as a first-line antimicrobial in all countries (n=11). Many of the second-line and alternative treatments seemed suboptimal for empirical treatment. Regionally manufactured antimicrobials were predominantly used and easily available, and some may be of suboptimal quality. This generates effective prerequisites for emergence, and rapid spread of gonococcal AMR and gonorrhoea. CONCLUSION: Ceftriaxone was first-line antimicrobial in all the 11 East European countries, which is an appropriate choice also in a global perspective. However, the adherence, especially among private physicians, to these public sector recommendations is questionable. Implementation of national and international gonococcal AMR surveillance in this region is crucial; to provide evidence-based data for regular and timely updating of treatment guidelines, to identify emerging resistance, and to assist in the prevention, control and containment of gonococcal AMR and gonorrhoea.

Chlamydia trachomatis serovar distributions in Russian men and women: a comparison with Dutch serovar distributions.

The data on serovar distributions of Chlamydia trachomatis - the most diagnosed sexually transmitted infection (STI) worldwide - are important for epidemiologic purposes and transmission studies but are completely lacking in Russia. The aim of the current study is to determine the serogroup and serovar distributions in Russian men and women and compare these data with Dutch serogroup and serovar distributions. In Russian men and women, serogroup B was the most prevalent (46%), followed by the intermediate serogroup (I group; 33%) and serogroup C (21%). The distribution was comparable between men and women. The serogroup distribution was similar to the previously published distribution in Dutch cohorts. However, on a serovar level statistically very significant differences were observed, reaching up to P < 0.0001. The serovars B and G/Ga had higher prevalences compared with the reported Dutch prevalences, while serovars F, H, I/Ia, J and K had lower prevalences compared with the Dutch studies. In conclusion, this is the first report of Russian C. trachomatis serovar/serogroup distributions. Serogroup B is the most prevalent, followed by serogroup I and serogroup C with no statistical differences on the serogroup level. However, significant differences between Russia and the Netherlands were observed in the distribution of C. trachomatis serovars.

Evaluation of six nucleic acid amplification tests used for diagnosis of Neisseria gonorrhoeae in Russia compared with an international strictly validated real-time porA pseudogene polymerase chain reaction.

BACKGROUND: In Russia, laboratory diagnosis of gonorrhoea has been mainly based on microscopy only and, in some settings, relatively rare suboptimal culturing. In recent years, Russian developed and manufactured nucleic acid amplification tests (NAAT) have been implemented for routine diagnosis of Neisseria gonorrhoeae. However, these NAATs have never been validated to any international well-recognized diagnostic NAAT. OBJECTIVE: This study aims to evaluate the performance characteristics of six Russian NAATs for N. gonorrhoeae diagnostics. MATERIALS AND METHODS: In total, 496 symptomatic patients were included. Five polymerase chain reaction (PCR) assays and one real-time nucleic acid sequence based amplification (NASBA) assay, developed by three Russian companies, were evaluated on urogenital samples, i.e. cervical and first voided urine (FVU) samples from females (n = 319), urethral and FVU samples from males (n = 127), and extragenital samples, i.e. rectal and pharyngeal samples, from 50 additional female patients with suspicion of gonorrhoea. As reference method, an international strictly validated real-time porA pseudogene PCR was applied. RESULTS: The prevalence of N. gonorrhoeae was 2.7% and 16% among the patients providing urogenital and extragenital samples, respectively. The Russian NAATs and the reference method displayed high level of concordance (99.4-100%). The sensitivities, specificities, positive predictive values and negative predictive values of the Russian tests in different specimens were 66.7-100%, 100%, 100%, and 99.4-100%, respectively. CONCLUSIONS: Russian N. gonorrhoeae diagnostic NAATs comprise relatively good performance characteristics. However, larger studies are crucial and, beneficially, the Russian assays should also be evaluated to other international highly sensitive and specific, and ideally Food and Drug Administration approved, NAATs such as Aptima Combo 2 (Gen-Probe).

First evaluation of polymerase chain reaction assays used for diagnosis of Mycoplasma genitalium in Russia.

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.

First evaluation of six nucleic acid amplification tests widely used in the diagnosis of Chlamydia trachomatis in Russia.

BACKGROUND: In Russia, nationally developed nucleic acid amplification tests (NAATs), which have never been validated to international commercially available NAATs, are mainly used in the diagnosis of Chlamydia trachomatis infection. OBJECTIVE: To evaluate the performance characteristics of six NAATs widely used to diagnose C. trachomatis infection in Russia. MATERIALS AND METHODS: In total, 446 consecutive symptomatic patients (319 females and 127 males) were included. Five polymerase chain reaction (PCR) assays and one real-time nucleic acid sequence-based amplification (NASBA) assay were evaluated on cervical and vaginal samples from females and on urethral and first voided urine samples from males. As reference methods, the Cobas Amplicor PCR, as the main 'gold standard' method, and LightMix 480HT PCR were used. RESULTS: The overall prevalence of C. trachomatis infection was 12.6%. The Russian NAATs and the reference methods displayed a high level of concordance (97.9% to 99.2%). In comparison with the reference methods, the sensitivities, specificities, positive predictive values and negative predictive values of the Russian tests in different specimens ranged from 86.1% to 100%, 99.1% to 100%, 92.3% to 100% and 98.2% to 100%, respectively. CONCLUSIONS: According to the reference methods, C. trachomatis NAATs developed and used in Russia have relatively good performance characteristics for both invasive and non-invasive samples. However, larger studies that include symptomatic and asymptomatic patients as well as genital and extra-genital samples, and in comparison with other internationally well-recognized, validated, and ideally Food and Drug Administration-approved C. trachomatis NAATs performed strictly according to the manufacturer's instructions, need to be conducted.