Objectives: Application of SYBR Green real-time PCR and bacterial isolation methods to detect Streptococcus agalactiae (GBS) carriage in women at 35-37 weeks’ gestation. Patients and Method: Use of SYBR Green real-time PCR and bacterial isolation methods to detect GBS in 116 women at 35 - 37 weeks’ gestation. Results: The rate of carrier of GBS in women at 35 - 37 weeks gestation was 9.5% (11 pregnant women), in which real-time PCR method was positive in all positive GBS women, while bacterial culture method was positive at 6% (7 pregnant women). These methods (culture and real-time PCR) had substantial agreement with the value of Kappa is 0,77. Checking for the targeted sequence amplification of real-time PCR by the curve of melting temperature and agarose electrophoresis of amplified product, the real-time PCR product was correctly targeted at the expected genetic sequence. Conclusion: SYBR Green real-time PCR method for detecting GBS in pregnant women is useful, low-cost and easy for performing. Therefore, it is suitable for detecting GBS in diagnostic laboratories where real-time PCRs are available. Key words: Streptococcus agalactiae (GBS), real-time PCR, pregnant woman
Background: The Nested IS6110 PCR is used for detecting tuberculosis, however IS6110 sequence is not present in the genome of all strains of M.tuberculosis, the result may be false negative. The gene coding 16S ribosome always contains a short sequence specific to M. tuberculosis complex. Objects: Performance of the 16S Real-time PCR to detect M. tuberculosis and combining to the nested IS6110 PCR to determine the rate of Mtb strains without IS6110 from clinical samples. Materials and method: Performance of 16S rDNA PCR by commercial kit of Viet A Inc. for all 480 samples, the samples which were positive with the 16S rDNA PCR were retested in IS6110 PCR assay by in-house kit. Results: The Realtime 16S rDNA PCR detected 258 cases (53.8%) of tuberculosis. There were 3 (1.2 %) M. tuberculosis strains which do not harbor IS6110 sequence in genome. Conclusion: The IS6110 nested PCR can be applied more widely than the 16S rDNA realtime PCR. In case of using IS6110 PCR assay, results may show a low proportion of false negative. Combining 16S rDNA PCR with the IS6110 based PCR allowed detection of deletion of IS6110 sequence in M. tuberculosis isolates.
Backround: Tuberculosis remains a major cause of morbidity and mortality in many countries and a significant public health problem worldwide. If TB is detected early and properly treated, the patients quickly become non-infectious and are eventually cured. Objective: Using MODS assay for detecting rapidly drug resistant and multidrug resistant M. tuberculosis. Methods: The MODS assay for the detection of tuberculosis and drug-resistant tuberculosis, directly from 252 sputum samples from suspected tuberculosis patients or untreated or ≤ 2 weeks treated tuberculosis patients. Culturing and doing susceptibility test by MODS assay (Issoniazid: 0.1 µg/ml; rifampicin: 1 µg/ml, Streptomycin: 2 µg/ml và Ethambutol: 2.5 µg/ml). Realtime PCR 16S was performed for the MODS culture-positive samples before 7 days and the samples with MODS culture-negative but AFB-positive. Results: M.tuberculosis was detected in 153 samples (60.7%) and 46 (30.1%) were antibiotic resistant. One drug resistance was present in 30 strains (19.6%): 18 for RIF, 6 for INH and 3 for STR and EBM. Multidrug resistant M.tuberculosis as defined by WHO (resistant to RIF and INH) was observed in 13 strains. There were additional 14 strains showing resistance to two or more drugs. Conclusion: The MODS assay is a rapid, direct method for simultaneous culture detection and drug susceptibility of M. tuberculosis, can be used as a routine procedure.
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