Nucleic Acid Amplification Tests (NAATs) for Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (GC) have become routine. These methods are validated for use with urogenital samples and have a faster turn-around-time for results with automation. Testing of non-validated samples is common which raises concerns about assay performance. In Australia, the 2005 Public Health Laboratory Network (PHLN) guideline recommends repeat testing of all initial positive GC NAAT results with a suitable alternate NAAT assay. In this study, results from > 70,000 patient samples from 2 laboratories were reviewed in accordance with the PHLN recommendations. The laboratories used the APTIMA Combo 2 (AC2) and APTIMA Neisseria gonorrhoeae (AGC) assays with the PANTHER instrument.72,253 AC2 results were available for analysis which included 1,174 (1.68%) initially AC2 reactive (positive or equivocal) samples which also had results for the AGC assay used as the confirmatory assay. For the reactive samples, the agreement between the AC2 and AGC assays occurred in 97.19% of samples; 1135 were positive in both assays and 6 were equivocal in both assays. Sample types tested included those with manufacturer's validation claims; urine, ThinPrep, vaginal, endocervical and urethral swabs and non-validated samples including throat, rectal, eye and joint fluids. Samples from throat swabs showed the highest numbers of discordant results followed by rectal swab samples. The percentage agreement of results obtained from all sample types was excellent, with an overall PPV of > 99%.Without confirmatory testing, false positive results would have been reported for 13 samples representing 0.02% of all samples tested. This study demonstrates that the initial AC2 results can be accepted with high confidence. Background Bacterial vaginosis (BV) is the most common cause of vaginal discharge in women. Since BV is associated with significant morbidity, accurate tools for diagnosing the disease are important. The Amsel criteria (AC) and Nugent score (NS) are currently used for BV diagnosis. Recently, a number of PCR-based tests providing objective, sensitive and specific BV detection have been described. This study aimed to evaluate a newly developed BV test based on quantitative detection of Gardnerella vaginalis, Atopobium vaginae, Lactobacillus spp. and total Bacteria using multiplex PCR. Methods PCR criteria were elaborated based on the relative counts of the targeted bacteria to classify vaginal microflora as BV, no BV or intermediate. Vaginal samples for the test evaluation were obtained from 274 patients addressing a gynaecologist for routine examination. All participants were of reproductive age, not pregnant and not menstruating at the time of enrollment. BV was diagnosed using the AC and NS. Results According to the NS results, 66 patients were BV positive, 156 were BV negative, and 52 were classified as intermediate. All patients positive by the NS were positive by the AC, and all patients negative by the NS were negative by the AC. Among 66 BV posi...

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.

The article presents literature review on the problem of bacterial vaginosis, describes current methods used for bacterial vaginosis diagnosis. Especial attention is spared for peculiarities vaginal microbiota at physiological microbiocenosis and with bacterial vaginosis.

ВведениеБактериальный вагиноз (БВ) является од-ной из важных проблем мирового здравоохра-нения, так как увеличивает риск инфицирова-ния ИППП и ВИЧ [6,15,24]. Он ассоциирован с нарушениями репродуктивного здоровья женщины, неудачными попытками ЭКО и аку-шерскими осложнениями, в том числе прежде-временными родами [10,12,14,22].Несколько лет тому назад многие авторы утверждали, что лишь Gardnerella vaginalis яв-ляется основным возбудителем БВ, существо-вал даже термин «гарднереллез». Позднее это утверждение было опровергнуто, так как этот микроорганизм стали выделять из влагалища здоровых женщин. Спектр микробных попу-ляций, ассоциированных с бактериальным ва-гинозом, был значительно расширен с началом применения молекулярных методов диагно-стики, которые обладают неоспоримым пре-имуществом по сравнению с культуральными, вследствие того что подавляющее большинство видов микроорганизмов трудно культивируют-ся или не культивируется в лабораторных усло-виях. В настоящее время общепризнанно, что ни один из этих микроорганизмов не является возбудителем бактериального вагиноза, а так как этиология БВ остается неясной, то и при-чины частых рецидивов -у 15-30 % женщин в течение 30-90 дней и в течение 9 месяцев у 70 % -остаются неизвестными [5,13,19].Несмотря на предложенный в 1993 г. Nugent и др. микроскопический метод [16], диагности-ка бактериального вагиноза основана преиму-

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.