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通过测量结核分枝杆菌三磷酸腺苷进行结核分枝杆菌药敏试验

[Antimicrobial susceptibility testing for Mycobacterium tuberculosis by measuring mycobacterial adenosine triphosphate].

作者信息

Yamazaki T, Sato N, Yamashita K, Okazawa Y, Miyao H, Tanno K

机构信息

Department of Bacteriology, National Institute of Infectious Diseases, Tokyo.

出版信息

Rinsho Byori. 1999 Feb;47(2):170-5.

Abstract

The antimicrobial susceptibility test for Mycobacterium tuberculosis H37Rv and 43 clinical isolates was performed using a bioluminescence assay by measuring the content of adenosine triphosphate (ATP) derived from mycobacteria. The drugs tested were isoniazid (INH), rifampicin (RFP), ethambutol (EB), streptomycin (SM), and kanamycin (KM). The ATP contents of M. tuberculosis incubated in the Middle-brook 7H9 broth medium containing antituberculous agents were measured at days of 0, 1, 3, 5, 7 and 10. A reduction of ATP content, indicating growth inhibition, was observed in susceptible strains within 5 to 10 days of incubation. Optimal concentrations to distinguish between susceptible and resistant strains were determined as being INH 0.20, RFP 0.50, EB 5.0, SM 4.0, KM 6.0 g/ml. The agreements of ATP method (evaluated at 10 days) with Vite Spectrum and MIC determinations were 81.4% and 100%, respectively. Susceptibilities to most drugs, except for EB, could be determined within 7 days. This method is simple, rapid, nonradiometric, and can be used for drug susceptibility.

摘要

采用生物发光法,通过测量结核分枝杆菌产生的三磷酸腺苷(ATP)含量,对结核分枝杆菌H37Rv及43株临床分离株进行了药敏试验。所检测的药物为异烟肼(INH)、利福平(RFP)、乙胺丁醇(EB)、链霉素(SM)和卡那霉素(KM)。在含有抗结核药物的Middle-brook 7H9肉汤培养基中培养的结核分枝杆菌,于第0、1、3、5、7和10天测量其ATP含量。在敏感菌株培养5至10天内观察到ATP含量降低,表明生长受到抑制。区分敏感和耐药菌株的最佳浓度确定为INH 0.20、RFP 0.50、EB 5.0、SM 4.0、KM 6.0μg/ml。ATP法(在第10天评估)与Vite Spectrum和MIC测定的一致性分别为81.4%和100%。除EB外,大多数药物的药敏情况可在7天内确定。该方法简单、快速、非放射性,可用于药敏试验。

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