Choi Seong Won, Maiga Mamoudou, Maiga Mariama C, Atudorei Viorel, Sharp Zachary D, Bishai William R, Timmins Graham S
Dept. Pharmaceutical Sciences, College of Pharmacy, University of New Mexico, Albuquerque NM 87131 USA.
Center for Tuberculosis Research, Department of Medicine, Johns Hopkins University, Baltimore MD 21231 USA.
Nat Commun. 2014 Sep 23;5:4989. doi: 10.1038/ncomms5989.
There is urgent need for rapid, point-of-care diagnostic tools for tuberculosis (TB) and drug sensitivity. Current methods based on in vitro growth take weeks, while DNA amplification can neither differentiate live from dead organisms nor determine phenotypic drug resistance. Here we show the development and evaluation of a rapid breath test for isoniazid (INH)-sensitive TB based on detection of labelled N2 gas formed specifically from labelled INH by mycobacterial KatG enzyme. In vitro data show that the assay is specific, dependent on mycobacterial abundance and discriminates between INH-sensitive and INH-resistant (S315T mutant KatG) TB. In vivo, the assay is rapid with maximal detection of (15)N2 in exhaled breath of infected rabbits within 5-10 min. No increase in (15)N2 is detected in uninfected animals, and the increases in (15)N2 are dependent on infection dose. This test may allow rapid detection of INH-sensitive TB.
迫切需要用于结核病（TB）和药物敏感性的快速即时诊断工具。目前基于体外培养的方法需要数周时间，而DNA扩增既无法区分活菌和死菌，也无法确定表型耐药性。在此，我们展示了一种基于检测由分枝杆菌KatG酶从标记异烟肼（INH）特异性生成的标记N2气体的快速呼气试验，用于检测对异烟肼敏感的结核病。体外数据表明，该检测具有特异性，依赖于分枝杆菌丰度，并且能够区分对异烟肼敏感和耐药（S315T突变型KatG）的结核病。在体内，该检测速度很快，感染兔子呼出的气体中在5-10分钟内可最大程度地检测到（15）N2。未感染动物中未检测到（15）N2增加，并且（15）N2的增加取决于感染剂量。该检测可能有助于快速检测对异烟肼敏感的结核病。
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