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用于分枝杆菌快速药敏试验的生物发光测定法的评估

Evaluation of a bioluminescence assay for rapid antimicrobial susceptibility testing of mycobacteria.

作者信息

Beckers B, Lang H R, Schimke D, Lammers A

出版信息

Eur J Clin Microbiol. 1985 Dec;4(6):556-61. doi: 10.1007/BF02013394.

Abstract

The antimicrobial susceptibility of Mycobacterium tuberculosis H37Rv and 17 clinical isolates of the same species was determined using a bioluminescence assay to measure adenosine triphosphate (ATP) produced by bacteria in the broth dilution test performed in Dubos-Tween-albumin broth. The results were compared with those obtained with the standard dilution proportional method performed on Löwenstein-Jensen medium. The drugs tested were streptomycin, rifampin, ethambutol and isoniazid. There was high correlation between bacterial count and ATP concentration in a liquid culture of Mycobacterium tuberculosis H37Rv (r = 0.993); when several cultures of the same strain were assayed after different incubation periods the correlation was somewhat lower (r = 0.846). Addition of tuberculostatic drugs in inhibitory concentrations resulted in a reduction in the ATP concentration as compared to control assays without drugs. Results of susceptibility tests performed by the two methods agreed in 66 out of 68 assays. Differences in the inhibitory concentrations determined by the two methods were due to differences in the methods. Whereas the standard method requires 3 to 4 weeks of incubation, results of the bioluminescence assay were available after five days of incubation. Automation of the assay for routine use is possible.

摘要

采用生物发光测定法,在杜博斯 - 吐温 - 白蛋白肉汤中进行肉汤稀释试验,测定结核分枝杆菌H37Rv及17株同物种临床分离株的抗菌药敏性,以测量细菌产生的三磷酸腺苷(ATP)。将结果与在罗 - 琴培养基上采用标准稀释比例法获得的结果进行比较。所测试的药物为链霉素、利福平、乙胺丁醇和异烟肼。在结核分枝杆菌H37Rv的液体培养物中,细菌计数与ATP浓度之间存在高度相关性(r = 0.993);当在不同培养时间后对同一菌株的多个培养物进行测定时,相关性略低(r = 0.846)。与无药物的对照试验相比,添加抑制浓度的抑菌药物会导致ATP浓度降低。两种方法进行的药敏试验结果在68次试验中有66次一致。两种方法测定的抑制浓度差异是由于方法不同所致。标准方法需要3至4周的培养时间,而生物发光测定法在培养五天后即可获得结果。该测定法可实现自动化以便常规使用。

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