ABSTRACT
Transcription profiles of genes of local immune response were determined in the vagina of women with bacterial vaginosis, aerobic vaginitis, and vulvovaginal candidosis for detection of the most specific immune markers for these vaginal infections. Laboratory diagnosis of the vaginal infections was performed microscopically; the inflammatory reaction in the vagina (leukorrhea) was defined as the presence of >10 white blood cells per field of view. Transcription profiles of IL1b, IL10, IL18, TNFα, TLR4, GATA3, and CD68 were determined using reverse-transcription quantitative real-time PCR. The strongest predictors of aerobic vaginitis were increased levels of IL1b and IL10 mRNA. Bacterial vaginosis was strongly associated with reduced levels of IL18 and GATA3 mRNA. Increased levels of IL1b and TLR4 transcripts showed significant discriminatory power for vulvovaginal candidosis and leukorrhea. The results of this study suggest differential expression of local immune response genes in the vagina of women with different vaginal infections. Detection of specific immune markers in the vagina using reverse-transcriptase PCR could supplement PCR detection of abnormal vaginal microflora for the diagnosis of vaginal infections.
Subject(s)
Adaptive Immunity/genetics , Vagina/immunology , Vagina/metabolism , Vaginosis, Bacterial/diagnosis , Adult , Biomarkers/analysis , Biomarkers/metabolism , Candidiasis, Vulvovaginal/diagnosis , Candidiasis, Vulvovaginal/genetics , Candidiasis, Vulvovaginal/immunology , Candidiasis, Vulvovaginal/microbiology , Case-Control Studies , Female , Gene Expression Regulation , Humans , Inflammation/genetics , Inflammation/immunology , Inflammation/metabolism , Interleukin-10/genetics , Interleukin-10/metabolism , Interleukin-1beta/genetics , Interleukin-1beta/metabolism , Toll-Like Receptor 4/genetics , Toll-Like Receptor 4/metabolism , Tumor Necrosis Factor-alpha/genetics , Tumor Necrosis Factor-alpha/metabolism , Vagina/pathology , Vaginosis, Bacterial/genetics , Vaginosis, Bacterial/immunology , Vaginosis, Bacterial/microbiologyABSTRACT
Resistance of 14 clinical isolates of C. trachomatis to fluoroquinolones, i.e. of ciprofloxacin, pefloxaxin and ofloxacin, was assayed. Three isolates with a high resistance degree to all 3 drugs (MIC equal or above 64 microg/ml) were detected. MIC was found to be equal to or below 4 microg/ml for 3 isolates. The remaining isolates had an intermediate resistance level. The nucleotide sequence was established for the Quinolone-Resistance Determining Region (QRDR) genes coding the DNA-gyrase subunit A (gyrA) and DNA-topoisomerase IV subunit C (parC) as well as for the 3'-region of ygeD coding, presumably, the efflux protein. In none of the isolates, the gyrA and gyrC QRDR differed from the corresponding regions in the published C. trachomatis genome sequence. Several silent mutations and mutations resulting in amino acid substitutions were observed in the ygeD 3' region of 2 isolates resistant to high FQ concentrations and in 1 isolate with the intermediate resistance level.
Subject(s)
Anti-Bacterial Agents/pharmacology , Chlamydia trachomatis/genetics , DNA Gyrase/genetics , DNA Topoisomerase IV/genetics , Fluoroquinolones/pharmacology , Chlamydia trachomatis/drug effects , Chlamydia trachomatis/isolation & purification , DNA Topoisomerases, Type II/genetics , Drug Resistance, Bacterial/genetics , Female , Humans , Microbial Sensitivity Tests , Point MutationABSTRACT
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.
