ABSTRACT
PURPOSE: To evaluate a real-time PCR-based technique to quantify bacteria associated with aerobic vaginitis (AV) as a potential test. METHODS: Vaginal samples from 100 women were tested by wet-mount microscopy, gram stain and quantitative real-time PCR targeting Enterobacteriacea, Staphylococcus spp., Streptococcus spp., Enterococcus spp., Escherichia coli, Streptococcus agalactiae, S. aureus; Lactobacillus spp. AV diagnosis obtained by wet-mount microscopy was used as reference. RESULTS: Some level of AV was diagnosed in 23 (23.7 %) cases. Various concentrations of Enterobacteriacea, Staphylococcus spp., Streptococcus spp. were detected an all patients. Enterococcus spp. were detected in 76 (78.3 %) cases. Summarized concentrations of aerobes were tenfold higher in AV-positive compared to AV-negative cases [7.30lg vs 6.06lg (p = 0.02)]. Concentrations of aerobes in severe, moderate and light AV cases did not vary significantly (p = 0.14). Concentration of lactobacilli was 1000-fold lower in AV-positive cases compared to normal cases (5.3lg vs 8.3lg, p < 0.0001). Streptococcus spp. dominated in the majority of AV-positive cases [19/22 (86.4 %) samples]. The relation of high loads of aerobes to the low numbers of Lactobacilli are a reliable marker for the presence of AV and could substitute microscopy as a test. CONCLUSIONS: PCR may be a good standardized substitution for AV diagnosis in settings where well-trained microscopists are lacking.
Subject(s)
Bacteria, Aerobic/genetics , Microscopy/methods , Real-Time Polymerase Chain Reaction/methods , Vagina/microbiology , Vaginosis, Bacterial/diagnosis , Adult , Bacteria, Aerobic/isolation & purification , Female , Humans , Middle Aged , Pregnancy , Russia , Sensitivity and Specificity , Vaginitis/diagnosis , Vaginitis/genetics , Vaginitis/microbiology , Vaginosis, Bacterial/genetics , Vaginosis, Bacterial/microbiologyABSTRACT
The technique is developed including corresponding kit of reagents "AmpliSens Florocenose/Candida-FL", to be applied in laboratory diagnostic of vuvlvovaginal candidiasis. The technique is based on polymerase chain reaction in real-time and permits identifying and quantitatively estimate in biological material content of five most prevalent agents of candidiasis: Candida albicans, C. glabrata, C. krusei, C. paprapsilosis, C. tropicalis. The experimental determination of analytical characteristics of developed technique was carried out. The analytical sensitivity amounted to 100 GE/mL (genome equivalent in 1 mL), linear range -from 200 to 2 x 10 7 GE/mL for every specie of detected Candida. The comparison of quantitative results of detection of content of Candida spp using the developed technique with results of inoculation demonstrated their mutual matching. In logarithm presentation the relation coefficient of lg (GE/mL) to lg (CFU/mL) amounted to 1.2. The correlation of results characterized by coefficient R 2 equal 0.76.