The Mycobacterium tuberculosis complex (MTBC) is composed of several genetically related and pathogenic mycobacterial species, including M. tuberculosis, M. bovis and M.africanum et al. In our previous study, we found that M. bovis strains had a unique SNP located in position 1055 in the sequence of the pstS1 gene in which a T was substituted by a C. In this study, specific primers and MGB probes were designed according to the mutation in PstS1 gene, and a sensitive, specific and rapid real-time PCR assay for M. bovis was established. Then the assay was used to detect M. bovis in simulation samples. The minimum detectable concentration is 10¹ copies for M. bovis DNA. The standard curve showed correlation coefficient between threshold cycle and PstS1 gene fragment copy number was 0.997 and slope is −3.144. The minimum detectable concentration is 10¹ cells/ml for simulation sample. In addition, M.bovis strain 93006, 14 clinical BCG stains and 7 clinical M.bovis strain showed positive while the other strains showed negative results, which proved good specificity. This assay had high sensitivity and specificity for identification of M. bovis from the simulation specimens. The assay can be applied for epidemiological and ecological surveillance of M. bovis strains.

在结核分枝杆菌复合（MTBC）是由几个基因相关的和致病分枝杆菌物种，包括结核分枝杆菌，牛分枝杆菌和M.africanum等。在我们以前的研究中，我们发现牛分枝杆菌菌株在pstS1基因的序列中第1055位有一个独特的SNP，其中T被C取代。在这项研究中，根据引物设计了特异性引物和MGB探针PstS1基因的突变，建立了一种针对牛分枝杆菌的灵敏，特异和快速的实时PCR检测方法。然后该测定法用于检测模拟样品中的牛分枝杆菌。最低可检测浓度为10 ¹牛分枝杆菌DNA的拷贝。标准曲线显示阈值循环与PstS1基因片段拷贝数之间的相关系数为0.997，斜率为-3.144。 模拟样品的最低可检测浓度为10 ¹细胞/ ml。另外，牛分枝杆菌菌株93006，14临床BCG污渍和7临床牛分枝杆菌菌株显示阳性，而其它菌株显示阴性结果，这证明了很好的特异性。该测定对于从模拟样品中鉴定牛分枝杆菌具有高灵敏度和特异性。该测定法可用于牛分枝杆菌菌株的流行病学和生态学监测。