Detection of Rifampicin Resistance in Mycobacterium tuberculosis by Using Middlebrook 7H9 Broth Medium with 2,3-Diphenyl-5-Thienyl-(2)-Tetrazolium Chloride

S. Lee, Kyung Jun Kim, Chulhun L. Chang
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Abstract

Background: A simple and cost-effective method is needed for the detection of rifampicin resistance in Mycobacterium tuberculosis in resource-limited settings. We suggest a broth medium-based method using 2,3-diphenyl-5-thienyl-(2)-tetrazolium chloride (STC) for detection of rifampin resistance of tubercle bacilli within a reasonable time frame. Methods: The type strain ( M. tuberculosis H37Rv) and 45 cultured clinical strains of M. tuberculosis (35 rifampin-susceptible and 10 rifampin-resistant) were used. Phenotypes of rifampicin resistance were tested by the Korea Institute of Tuberculosis, and con-firmed by GenoType MTBDRplus (Hain Lifescience, Germany). Susceptibility tests were performed using STC-containing OADC-enriched broth (BD, USA). Results: All tests were finished in 3 to 6 days. The same results were obtained with the standard and current methods for all 45 clinical isolates (100% sensitivity and specificity for resistance detection). Conclusion: The current method using STC is a good alternative for detecting M. tuberculosis rifampin resistance in a cost-effective and timely fashion, which is particularly important in resource-limited settings. (Ann Clin Microbiol 2018;21:47-50)
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用含2,3-二苯基-5-噻吩基-(2)-四唑氯化铵的Middlebrook 7H9肉汤培养基检测结核分枝杆菌对利福平的耐药性
背景:在资源有限的情况下,需要一种简单且具有成本效益的方法来检测结核分枝杆菌中的利福平耐药性。我们提出了一种基于肉汤培养基的方法,使用2,3-二苯基-5-噻吩基-(2)-氯化四氮唑(STC)在合理的时间范围内检测结核杆菌的利福平耐药性。方法:采用结核分枝杆菌H37Rv型菌株和45株临床培养的结核分枝杆菌(35株对利福平敏感,10株对利福平耐药)。利福平耐药性表型由韩国结核病研究所测试,并由基因型MTBDRplus(德国海恩生命科学公司)确认。使用含有STC的OADC富集肉汤(BD,USA)进行易感性测试。结果:所有试验均在3-6天内完成。所有45个临床分离株的标准和现行方法都获得了相同的结果(耐药性检测的敏感性和特异性为100%)。结论:目前使用STC的方法是一种经济高效、及时检测结核分枝杆菌利福平耐药性的好方法,这在资源有限的环境中尤为重要。(Ann Clin Microbiol 2018;21:47-50)
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