Eisenach K D, Cave M D, Bates J H, Crawford J T
Medical Research Service, McClellan Memorial Veterans Hospital, Little Rock, AR 72205.
J Infect Dis. 1990 May;161(5):977-81. doi: 10.1093/infdis/161.5.977.
A segment of DNA repeated in the chromosome of Mycobacterium tuberculosis was sequenced and used as a target for amplification using polymerase chain reaction (PCR). The sequences of the primers (5' to 3') were CCTGCGAGCGTAGGCGTCGG and CTCGTCCAGCGCCGCTTCGG, and a temperature of 68 degrees C was used for annealing the primers in the reaction. Amplification produced a 123-base-pair fragment with an internal SalI site. The specific PCR product was obtained with input DNA from 11 different strains of M. tuberculosis and Mycobacterium bovis and one strain of Mycobacterium simiae. No product was detected with DNA from 28 strains of the Mycobacterium avium complex, Mycobacterium scrofulaceum, Mycobacterium kansasii, Mycobacterium fortuitum, Mycobacterium chelonei, and Mycobacterium gordonae. The PCR product was detected by gel electrophoresis after 30 cycles using 1 fg of input DNA. Amplification of this sequence may provide the basis for an assay to detect M. tuberculosis directly in clinical material.
对结核分枝杆菌染色体中重复的一段DNA进行了测序，并将其用作聚合酶链反应（PCR）扩增的靶标。引物（5'至3'）序列为CCTGCGAGCGTAGGCGTCGG和CTCGTCCAGCGCCGCTTCGG，反应中引物退火温度为68℃。扩增产生了一个123个碱基对的片段，该片段带有一个内部SalI位点。从11种不同的结核分枝杆菌和牛分枝杆菌菌株以及1种猿分枝杆菌菌株的输入DNA中获得了特异性PCR产物。用鸟分枝杆菌复合群、瘰疬分枝杆菌、堪萨斯分枝杆菌、偶然分枝杆菌、龟分枝杆菌和戈登分枝杆菌的28个菌株的DNA未检测到产物。使用1 fg输入DNA经过30个循环后，通过凝胶电泳检测到PCR产物。该序列的扩增可为直接在临床材料中检测结核分枝杆菌的检测方法提供基础。
