用于诊断肺结核的噬菌体复制检测方法的开发。
Development of a bacteriophage phage replication assay for diagnosis of pulmonary tuberculosis.
作者信息
McNerney Ruth, Kambashi Bupe S, Kinkese Juliana, Tembwe Ruth, Godfrey-Faussett Peter
机构信息
London School of Hygiene and Tropical Medicine, London WC1E 7HT, United Kingdom.
出版信息
J Clin Microbiol. 2004 May;42(5):2115-20. doi: 10.1128/JCM.42.5.2115-2120.2004.
Successful infection and replication of bacteriophages is indicative of the presence of viable bacteria. We describe here the development of a bacteriophage replication assay for the detection of Mycobacterium tuberculosis by using mycobacteriophage D29. Optimization of phage inoculate and incubation times allowed highly sensitive detection of M. bovis BCG. Fewer than 10 CFU (100 CFU/ml) were detected. No false-positive results were observed in negative samples. Application of the assay to 496 sputum specimens in the National Reference Laboratory of Zambia produced sensitivity, specificity, and positive and negative predictive values of 44.1, 92.6, 82.2, and 67.5%, respectively, compared to culture on Lowenstein-Jensen medium. The equivalent corresponding results for direct fluorescent smear microscopy were 42.3, 96.8, 91.2, and 67.6%. The small increase in sensitivity over that of direct microscopy does not justify the introduction of this technique for routine diagnosis of pulmonary tuberculosis at this time.
噬菌体的成功感染和复制表明存在活细菌。我们在此描述了一种利用分枝杆菌噬菌体D29检测结核分枝杆菌的噬菌体复制检测方法的开发。对噬菌体接种量和孵育时间的优化使得能够高度灵敏地检测牛分枝杆菌卡介苗。检测到的菌落形成单位少于10个（每毫升100个菌落形成单位）。在阴性样本中未观察到假阳性结果。与在罗-琴培养基上培养相比，在赞比亚国家参考实验室将该检测方法应用于496份痰标本时，其敏感性、特异性、阳性预测值和阴性预测值分别为44.1%、92.6%、82.2%和67.5%。直接荧光涂片显微镜检查的相应等效结果为42.3%、96.8%、91.2%和67.6%。与直接显微镜检查相比，敏感性虽有小幅提高，但目前还不足以证明引入该技术用于肺结核的常规诊断是合理的。
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