First antimicrobial resistance data and genetic characteristics of Neisseria gonorrhoeae isolates from Estonia, 2009-2013.

Gonorrhoea is a sexually transmitted infection with major public health implications and Neisseria gonorrhoeae has developed resistance to all antimicrobials introduced for treatment. Enhanced surveillance of antimicrobial resistance in N. gonorrhoeae is crucial globally. This is the first internationally reported antimicrobial resistance data for N. gonorrhoeae from Estonia (44 isolates cultured in 2009-2013). A high prevalence of resistance was observed for azithromycin, ciprofloxacin and tetracycline. One and two isolates with resistance and decreased susceptibility to the last remaining first-line treatment option ceftriaxone, respectively, were identified. It is crucial to implement surveillance of gonococcal antimicrobial resistance (ideally also treatment failures) in Estonia.

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.

Guidelines for the laboratory diagnosis of genital herpes in eastern European countries.

These guidelines aim to provide comprehensive information about sexually transmitted herpes simplex virus (HSV) infection and its laboratory diagnosis in eastern European countries. They are primarily intended for professionals testing specimens from patients at a sexual healthcare clinic but may also be helpful for community-based screening programmes. In particular, the guidelines recommend: (i) either viral culture or validated and approved nucleic acid amplification tests (NAATs) as the tests of choice for symptomatic patients, which should be promoted for laboratory confirmation of HSV infection; (ii) if culture or NAATs are not available, antigen detection--a direct immunofluorescence test or enzyme immunoassay from samples from symptomatic patients--could be employed, but HSV type determination is of importance; (iii) only type-specific serology should be used for detecting asymptomatic individuals, testing pregnant women at risk of acquiring HSV infection close to delivery, men who have sex with men and people who are HIV positive; (iv) widespread screening for HSV antibodies should be discouraged; and (v) any non-validated diagnostic tests should be validated against a recommended, approved gold standard.

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.

The Russian gonococcal antimicrobial susceptibility programme (RU-GASP)--national resistance prevalence in 2007 and 2008, and trends during 2005-2008.

Antimicrobial resistance (AMR) in Neisseria gonorrhoeae is a major problem worldwide. In the former Soviet countries including Russia, the knowledge regarding AMR has been highly limited. However, in 2004 the Russian gonococcal antimicrobial susceptibility programme (RU-GASP) was initiated. The aims of this study were to examine and describe the prevalence of N. gonorrhoeae AMR in 2007 and 2008 in Russia, and reveal trends in the period from 2005 to 2008. Gonococcal isolates (660 in 2007 and 900 in 2008) from 36 surveillance sites were examined using agar dilution method. From 2005 to 2008, the proportion of isolates resistant to spectinomycin increased from 0% to 7.2%, and remained high for those resistant to ciprofloxacin (approximately 49%). The resistance to azithromycin was 2.3% and 0.4% in 2007 and 2008, respectively. All isolates between 2005 and 2008 were susceptible to ceftriaxone. In conclusion, the AMR of N. gonorrhoeae in Russia is high, as in most countries in the European Union, and ceftriaxone should be the first line for treatment. If there is no access to ceftriaxone or in the presence of severe beta-lactam antimicrobial allergy, spectinomycin should be used; however, the resistance to spectinomycin has increased. Regular, quality-assured national and international surveillance of AMR in N. gonorrhoeae is crucial globally for public health.

Guidelines for the laboratory diagnosis of trichomoniasis in East European countries.

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.

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.

Laboratory support for the diagnosis and surveillance of sexually transmitted infections (STIs) in Eastern Europe.

This report outlines the proceedings of the 4th Annual Meeting of the Eastern European Network for Sexual and Reproductive Health (EE SRH Network) [1,2], which took place at Uppsala University in Uppsala, Sweden between 30 May and 3 June, 2009.

Guidelines for the laboratory diagnosis of Chlamydia trachomatis infections in East European countries.

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.

Guidelines for the laboratory diagnosis of syphilis in East European countries.

The present guidelines aim to provide comprehensive and precise information regarding the laboratory diagnosis of the sexually transmitted infection (STI) syphilis in East European countries. These recommendations contain important information for laboratory staff working with 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.

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.

Implementation of a national electronic reporting system in Lithuania.

Electronic reporting systems improve the quality and timeliness of the surveillance of communicable diseases. The aim of this paper is to present the process of the implementation and introduction of an electronic reporting system for the surveillance of communicable diseases in Lithuania. The project which started in 2002 was performed in collaboration between Lithuania and Sweden and was facilitated by the parallel process of adapting the surveillance system to European Union (EU) standards. The Lotus-based software, SmittAdm, was acquired from the Department of Communicable Diseases Control and Prevention of Stockholm County in Sweden and adopted for Lithuania, resulting in the Lithuanian software, ULISAS. A major advantage of this program for Lithuania was the possibility to work offline. The project was initiated in the two largest counties in Lithuania where ULISAS had been installed and put in use by January 2005. The introduction was gradual, the national level was connected to the system during late 2005, and all remaining counties were included during 2006 and 2007. The reporting system remains to be evaluated concerning timeliness and completeness of the surveillance. Further development is needed, for example the inclusion of all physicians and laboratories and an alert system for outbreaks. The introduction of this case-based, timely electronic reporting system in Lithuania allows better reporting of data to the European Centre for Disease Prevention and Control (ECDC) and the World Health Organization (WHO) compared to the former reporting system with paper-based, aggregated data.

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.

Laboratory diagnostics for non-viral sexually transmitted infections in St. Petersburg, Russia: current situation and hallmarks for improvements.

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.

National surveillance of antimicrobial susceptibility in Neisseria gonorrhoeae in 2005-2006 and recommendations of first-line antimicrobial drugs for gonorrhoea treatment in Russia.

OBJECTIVES: To investigate comprehensively the antimicrobial susceptibility and resistance of Neisseria gonorrhoeae during 2005-2006 in a national survey and to recommend effective antimicrobial drugs for the treatment of gonorrhoea in Russia. METHODS: The susceptibility of N gonorrhoeae isolates, cultured mainly from consecutive gonorrhoea patients (n = 1030) during the period January 2005 to December 2006 in Russia, to penicillin G, ceftriaxone, ciprofloxacin, tetracycline and spectinomycin was analysed using the agar dilution method. Nitrocefin discs were used for beta-lactamase detection. RESULTS: All isolates were susceptible to ceftriaxone. During 2005 and 2006, however, 5%, 50%, 70% and 77% displayed intermediate susceptibility or resistance to spectinomycin, ciprofloxacin, tetracycline and penicillin G, respectively. Furthermore, 4% of the isolates were beta-lactamase producing during these years. The different federal districts of Russia displayed substantial heterogeneities with regard to the prevalence of gonorrhoea and antimicrobial resistance among N gonorrhoeae isolates. CONCLUSIONS: In Russia, penicillins, ciprofloxacin, or tetracycline should definitively not be used in the empirical treatment of gonorrhoea. The recommended first-line antimicrobial drug should be ceftriaxone. If ceftriaxone is not available, spectinomycin ought to be used. Increasing levels of intermediate susceptibility and resistance to spectinomycin have, however, been observed during recent years and, accordingly, great care and monitoring should be undertaken when using this agent. Continuous local, national and international surveillance of N gonorrhoeae antimicrobial susceptibility, in order to reveal the emergence of new resistance, to monitor changing patterns of susceptibility and to be able to update treatment recommendations on a regular basis, is crucial.