Tortoli E, Nanetti A, Piersimoni C, Cichero P, Farina C, Mucignat G, Scarparo C, Bartolini L, Valentini R, Nista D, Gesu G, Tosi C P, Crovatto M, Brusarosco G
Centro Regionale di Riferimento per la Diagnostica delle Micobatteriosi, Laboratorio di Microbiologia e Virologia, Ospedale di Careggi, Piastra del servizi, viale Morgagni 85, 50134 Florence, Italy.
J Clin Microbiol. 2001 Mar;39(3):1079-84. doi: 10.1128/JCM.39.3.1079-1084.2001.
A new DNA probe assay (INNO LiPA Mycobacteria; Innogenetics, Ghent, Belgium) for the simultaneous identification, by means of reverse hybridization and line-probe technology, of Mycobacterium tuberculosis complex, Mycobacterium kansasii, Mycobacterium xenopi, Mycobacterium gordonae, the species of the Mycobacterium avium complex (MAC), Mycobacterium scrofulaceum, and Mycobacterium chelonae was evaluated on a panel of 238 strains including, besides representatives of all the taxa identifiable by the system, a number of other mycobacteria, some of which are known to be problematic with the only other commercial DNA probe system (AccuProbe; Gen-Probe, San Diego, Calif.), and two nocardiae. The new kit, which includes a control probe reacting with the whole genus Mycobacterium, correctly identified 99.6% of the strains tested; the one discrepancy, which remained unresolved, concerned an isolate identified as MAC intermediate by INNO LiPA Mycobacteria and as Mycobacterium intracellulare by AccuProbe. In five cases, because of an imperfect checking of hybridization temperature, a very slight, nonspecific, line was visible which was no longer evident when the test was repeated. Two strains whose DNA failed amplification at the first attempt were regularly identified when the test was repeated. Interestingly, the novel kit dodged all the pitfalls presented by the strains giving anomalous reactions with AccuProbe. A unique feature of INNO LiPA Mycobacteria is its ability to recognize different subgroups within the species M. kansasii and M. chelonae, while the declared overlapping reactivity of probe 4 with some M. kansasii and Mycobacterium gastri organisms and of probe 9 with MAC, Mycobacterium haemophilum, and Mycobacterium malmoense, may furnish a useful aid for their identification. The turnaround time of the method is approximately 6 h, including a preliminary PCR amplification.
一种新的DNA探针检测方法（INNO LiPA分枝杆菌检测试剂盒；Innogenetics公司，比利时根特），通过反向杂交和线性探针技术，可同时鉴定结核分枝杆菌复合群、堪萨斯分枝杆菌、偶发分枝杆菌、戈登分枝杆菌、鸟分枝杆菌复合群（MAC）、瘰疬分枝杆菌和龟分枝杆菌。我们用238株菌株组成的菌株库对该方法进行了评估，除了该系统可鉴定的所有分类群的代表菌株外，还包括一些其他分枝杆菌，其中一些已知在用另一种商业DNA探针系统（AccuProbe；Gen-Probe公司，美国加利福尼亚州圣地亚哥）检测时存在问题，以及两株诺卡菌。该新试剂盒包含一个与整个分枝杆菌属反应的对照探针，正确鉴定了99.6%的测试菌株；唯一未解决的差异是，一株菌株被INNO LiPA分枝杆菌检测试剂盒鉴定为MAC中间型，而被AccuProbe鉴定为胞内分枝杆菌。在5个案例中，由于杂交温度检查不完善，出现了一条非常轻微的非特异性条带，重复测试时该条带不再明显。有两株菌株的DNA在首次尝试时扩增失败，但重复测试时能正常鉴定。有趣的是，该新型试剂盒避开了AccuProbe检测时出现异常反应的菌株所带来的所有陷阱。INNO LiPA分枝杆菌检测试剂盒的一个独特特点是能够识别堪萨斯分枝杆菌和龟分枝杆菌种内的不同亚群，而探针4与一些堪萨斯分枝杆菌和胃分枝杆菌的声明重叠反应性，以及探针9与MAC、嗜血性分枝杆菌和马尔默分枝杆菌的重叠反应性，可能有助于它们的鉴定。该方法的周转时间约为6小时，包括初步的PCR扩增。
