[A new multiplex allele-specific polymerase chain reaction assay for detection of rifampin-resistant Mycobacterium tuberculosis].
OBJECTIVE
To develop a new multiplex allele-specific polymerase chain reaction (MAS-PCR) assay to detect the main mutations in the rifampin resistance dependent region, which has been reported to account for the majority of clinic Mycobacterium tuberculosis resistant to rifampin.


METHODS
Based on the sequence of rpoB gene, three specific primers were designed for the MAS-PCR to detect the most common mutations in codons 531, 526, 516 of rpoB gene.


RESULTS
The purified DNA preparations of 91 clinical strains of Mycobacterium tuberculosis were used to optimize the PCR. The mutations in codon 531, 526, 516 were detected by the MAS-PCR. Compared with the results of direct sequencing of rpoB gene, no mutation was detected in the sensitive strains. For rifampin-resistant strains, the total sensitivity was 81.5% (66/81).


CONCLUSIONS
MAS-PCR is a new molecular method with a high sensitivity and specificity, which can be used to detect the 3 main mutations of rpoB gene rapidly and economically. It can be used in clinical laboratories to detect the rifampin-resistant strains of Mycobacterium tuberculosis.
Jin
Jia-lin
JL
Department of Infectious Diseases, Huashan Hospital, Fudan University, Shanghai 200040, China.
Zhang
Wen-hong
WH
Hu
Zhong-yi
ZY
Chen
Shu
S
Weng
Xin-hua
XH
D000483:Alleles; D000995:Antitubercular Agents / Q000494:pharmacology; D004269:DNA, Bacterial / Q000235:genetics; D024881:Drug Resistance, Bacterial; D005798:Genes, Bacterial; D008826:Microbial Sensitivity Tests / Q000379:methods; D009169:Mycobacterium tuberculosis / Q000187:drug effects* / Q000235:genetics / Q000302:isolation & purification*; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D012293:Rifampin / Q000494:pharmacology; D012680:Sensitivity and Specificity
D000995:Antitubercular Agents; D004269:DNA, Bacterial; D012293:Rifampin