Title: Evaluation of PCR-probe melting analysis assay for detection of rifampin-resistance in Mycobacterium tuberculosis clinical isolates
Abstract:Objective To evaluate the clinical application of PCR-probe melting analysis assay (PMAA) for detection of rifampin (RFP)-resistantce in Mycobacterium tuberculosis (MTB) clinical isolates. Methods Con...Objective To evaluate the clinical application of PCR-probe melting analysis assay (PMAA) for detection of rifampin (RFP)-resistantce in Mycobacterium tuberculosis (MTB) clinical isolates. Methods Conventional proportion method (PM) and PCR-PMAA were used to do RFP drug susceptibility test of 512 MTB clinical isolates. The rpoB genes of all clinical isolates were sequenced. Results Using PM as control, the sensitivity, specificity, positive predictive value,negative predictive value, coincidence rate and Youden index with PCR-PMAA of RFP-resistance test were 97.5%,96.2%,94.1%,98.4%,96.7% and 0.93 respectively. In 193 RFP-resistant clinical isolates detected by both PM and PCR-PMAA methods showed that, 174 had single codon mutations and 19 had double codon mutations in RFP resistance determining region of rpoB gene; 5 clinical isolates were RFP-resistant by PM method but RFP-sensitive by PCR-PMAA method had no mutations in rpoB genes; Among 12 clinical isolates of RFP-sensitive by PM method and RFP-resistant by PCR-PMAA method, 7 mutations were at 511 codon and 5 mutations were at 533 codon in rpoB gene. No mutations associated with resistance in rpoB gene were found in 302 RFP-sensitive clinical isolates by both PM and PCR-PMAA methods. The mutations in rpoB genes of MTB clinical isolates tested by PCR-PMAA method were identical with DNA sequencing. Conclusions PCR-PMAA method could effectively detect mutations in RFP-resistant genes of MTB, and it is sensitive, specific, simple, rapid and inexpensive for RFP-resistance determination of MTB. It possesses a good prospects in clinical application.Read More
Publication Year: 2012
Publication Date: 2012-01-01
Language: en
Type: article
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