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.
The sensitivity of 11 clinical strains of Chlamydia trachomatis to azithromycin, ofloxacin, doxycycline, and erythromycin was evaluated. The minimum inhibiting concentrations of all antibiotics for 90% strains, determined by PCR with reverse transcription of omp3B gene RNA (GenBank U68443) corresponded to, and those with reverse transcription of 16S rRNA gene RNA (GenBank X54451) far surpassed the minimum bactericidal concentrations for 90% strains determined by direct immunofluorescence with monoclonal antibodies to the major outer membrane protein.
The technique of pooling endocervical samples for PCR detection of C. trachomatis was developed and compared with individual testing. The efficiency of pooling strategy was evaluated for its accuracy and cost saving ability. Population prevalence based on pooled data was estimated. 1. 500 endocervical samples were tested individually and pooled by 5 (300 pools) and 10 (150 pools) specimens. The sensitivity and specificity of PCR was not affected by pooling either by 5 or by 10 samples. The estimated prevalence was 6. 1% (95% CI: 4,5-7,7) and 6,0% (95% CI: 4,3-7,) for pooling by 5 and 10, respectively. The prevalence of 6. 6% determined by individual testing (99 of 1. 500) was within 95% CI of the estimated prevalence for pooling by 5 and 10. The used pooling strategy has resulted in 53. 3 and 44. 0% cost savings, when endocervical samples were pooled by 5 and 10, respectively. Thus, pooling endocervical samples for detection of C. trachomatis is an accurate and cost saving approach for realization of large-scale studies.
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