监狱的电视剧:JCM：利用荧光噬菌体开发出更快速简便的检测结核病方法

结核分枝杆菌(Mycobacterium tuberculosis)，图片来自美国疾病防控中心公共健康图片库，Janice Haney Carr。

根据2012年1月25日发表在Journal of Clinical Microbiology期刊上的一篇研究论文，一个国际研究小组优化一种让结核分枝杆菌(Mycobacterium tuberculosis)在显微镜下发出明亮绿色荧光的策略。这种筛选活动性结核病(active tuberculosis)的新方法可能很快给人们提供一种更快度和更简便的检测方法。荧光噬菌体(fluorophage)是携带荧光报告基因的细菌病毒。研究人员利用该噬菌体去感染从病人肺部咳出的含有结核菌素(tuberculin)唾液中的分枝杆菌。几个小时之内，被感染的细菌表达报告基因而且发出的荧光水平足够高使得人们在显微镜下也能观察到。

以前有人试图构建类似地筛选结核病细菌的方法，但是遭遇到报告基因表达过低的难题。为了解决这种问题，研究人员首先从克隆能力较强的噬菌体开始研究，给它加入一个活性比较强的噬菌体启动子。这种方法产生的荧光信号是以前报到的100倍，而且分枝杆菌阳性的临床样品无需对细菌进行培养就可在显微镜下检测到。此外，这种技术还允许人们进行细菌耐药性筛查。在抗生素存在下，耐药性细菌继续发出荧光，而药物敏感性细菌因不能表达报告基而不能发出荧光。(生物谷：towersimper编译)

doi:10.1128/?JCM.06192-11
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2GFP10: A high-intensity fluorophage enables detection and rapid drug susceptibility testing of Mycobacterium tuberculosis directly from sputum samples

Paras Jain, Travis E. Hartman, Nell Eisenberg, Max R. O'Donnel, Jordan Kriakov, Karnishree Govender, Mantha Makume, David S. Thaler, Graham F. Hatfull, A. Willem Sturm, Michelle H. Larsen, Preshnie Moodley and William R. Jacobs Jr.

The diagnosis of active tuberculosis (TB) and rapid drug susceptibility testing (DST) at point of care remain critical obstacles to TB control. This report describes a high-intensity mycobacterial-specific-fluorophage (φ2GFP10) that for the first time allows direct visualization of Mycobacterium tuberculosis (Mtb) in clinical sputum samples. Engineered features distinguishing φ2GFP10 from previous reporter phages include an improved vector backbone with increased cloning capacity and superior expression of fluorescent reporter genes through use of an efficient phage promoter. φ2GFP10 produces a 100-fold increase in fluorescence-per-cell compared to existing reporter-phages. DST for isoniazid and oxofloxacin, carried out in cultured samples, was complete within 36 h. φ2GFP10 detected M. tuberculosis in clinical sputum samples collected from TB patients. DST for rifampicin and kanamycin from sputum samples yielded results after 12h of incubation with φ2GFP10. Fluorophage φ2GFP10 has potential for clinical development as a rapid, sensitive, and inexpensive point-of-care diagnostic tool for M. tuberculosis infection, and as a rapid DST.