Riska P F, Jacobs W R, Bloom B R, McKitrick J, Chan J
Division of Infectious Diseases, Montefiore Medical Center of the Albert Einstein College of Medicine, Bronx, New York 10461, USA.
J Clin Microbiol. 1997 Dec;35(12):3225-31. doi: 10.1128/jcm.35.12.3225-3231.1997.
We have previously described a luciferase reporter mycobacteriophage (LRP) assay that can detect Mycobacterium tuberculosis and characterize mycobacterial drug susceptibility patterns within 24 to 48 h in positive cultures. One drawback of this LRP protocol is the ability of the recombinant mycobacteriophage phAE40 to infect a variety of Mycobacterium species, thus limiting its specificity for the detection of M. tuberculosis. In this study, we have (i) explored the host range of phAE40, (ii) developed a modified LRP assay that exploits the selective inhibitory effect of the compound p-nitro-alpha-acetylamino-beta-hydroxy propiophenone (NAP) against members of the M. tuberculosis complex to differentiate between the tubercle bacillus and other mycobacterial species, and (iii) tested over 300 samples, including primary clinical isolates and drug-resistant strains of M. tuberculosis, demonstrating the ability of the NAP-modified LRP assay to identify M. tuberculosis complex organisms with high degrees of sensitivity and specificity.
我们之前描述了一种荧光素酶报告分枝杆菌噬菌体（LRP）检测方法，该方法可在阳性培养物中24至48小时内检测结核分枝杆菌并表征分枝杆菌药物敏感性模式。此LRP方案的一个缺点是重组分枝杆菌噬菌体phAE40能够感染多种分枝杆菌物种，从而限制了其检测结核分枝杆菌的特异性。在本研究中，我们（i）探索了phAE40的宿主范围，（ii）开发了一种改良的LRP检测方法，该方法利用化合物对硝基-α-乙酰氨基-β-羟基苯丙酮（NAP）对结核分枝杆菌复合群成员的选择性抑制作用来区分结核杆菌和其他分枝杆菌物种，并且（iii）检测了300多个样本，包括结核分枝杆菌的原发性临床分离株和耐药菌株，证明了NAP改良的LRP检测方法能够高度灵敏且特异地鉴定结核分枝杆菌复合群生物体。
