A New Genetic Assay for Rifampicin Susceptibility of <I>Mycobacterium tuberculosis</I>

KUSUNOKI Shinji, TAKEMURA Kiyoshi, TAKAGI Keiko, INAGAKI Yuji, ASAKA Jun-ichiro, EZAKI Takayuki, IGIMI Hirotsune

doi:10.11150/kansenshogakuzasshi1970.69.1342

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<I>Mycobacterium tuberculosis</I>のrifampicin感受性を知る新しい遺伝子学的試験法
Mycobacterium tuberculosisのrifampicin感受性を知る新しい遺伝子学的試験法〔英文〕
Mycobacterium tuberculosis ノ rifampicin
A New Genetic Assay for Rifampicin Susceptibility of Mycobacterium tuberculosis

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Abstract

A new method for testing rifampicin (RFP) susceptibility of Mycobacterium tuberculosis was developed. This method is based on detection of the internal sequence derived from 71-kDa heat shock protein mRNA in tubercle bacilli heat-treated in the presence of RFP. The target sequence was amplified by reverse transcription and PCR, followed by agarose gel electrophoretic analysis. No amplification occurred in one RFP-susceptible strain by exposure to 45°C for 45 min in Middlebrook 7H9 broth containng RFP (10μg/ml) after overnight incubation at 37°C. On the other hand, an amplified 275-bp product was obtained from the RFP-resistant strain MY-129. In a subsequent trial using 65 clinical isolates, this method defined their RFP susceptibility levels as well as the verification of the MICs obtained by the conventional agar dilution method, with the exception of one RFP susceptible strain. Thus, this method provides a rapid and practical system to determine RFP susceptibility in M. tuberculosis.

Journal

Kansenshogaku Zasshi

Kansenshogaku Zasshi 69 (12), 1342-1347, 1995

The Japanese Association for Infectious Diseases

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