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  • Title: [A new multiplex allele-specific polymerase chain reaction assay for detection of rifampin-resistant Mycobacterium tuberculosis].
    Author: Jin JL, Zhang WH, Hu ZY, Chen S, Weng XH.
    Journal: Zhonghua Jie He He Hu Xi Za Zhi; 2005 Sep; 28(9):623-5. PubMed ID: 16207432.
    Abstract:
    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.
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