Evaluation for rapid detection of rifampicin-resistant mycobacterium tuberculosis by polymerase chain reaction-single strand conformation polymorphism

Abstract

We evaluated usefulness of the rapid diagnostic method for detection of rifampicin (RFP)-resistant Mycobacterium tuberculosis, which was based on polymerase chain reaction. The MICs of RFP were measured for 38 clinical isolates of Mycobacterium tuberculosis which were suspected to be RFP-resistant organisms, and 12 strains were found to be resistant to RFP. The PCR primers used were the same as those reported by Telenti et al, which were targeting the RNA polymerase beta subunit gene (rpoB). We confirmed that this gene was possessed by all the strains tested. Eight strains out of the 12 strains with RFP-resistant phenotype were demonstrated to have a point mutation or some alterationin the rpoB gene on the basis of PCR-single strand conformation polymorphism (SSCP). Thus, the sensitivity of our method was calculated as 67%. In addition, we could not detect any alterations in the rpoB gene by all RFP-susceptible strains. These results indicated that rapid detection of the RFP-resistant Mycobacterium tuberculosis was possible directly from clinical specimens by using PCR-SSCP technique.