Das Samir, Das Suresh Chandra, Verma Rishendra
Division of Veterinary Public Health, Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, India.
Microbiol Immunol. 2007;51(2):231-4. doi: 10.1111/j.1348-0421.2007.tb03905.x.
The present investigation dealt with the identification of Mycobacterium tuberculosis and M. bovis by RD9 region and 500 bp fragment PCR assays. Eight M. tuberculosis and 5 M. bovis characterized and identified from 40 human sputum and 41 bovine lung specimens and 20 M. tuberculosis and 9 M. bovis strains maintained at Mycobacteria Laboratory, Indian Veterinary Research Institute were included in this study. In this way, 28 M. tuberculosis and 14 M. bovis strains and, for comparison and control purpose, M. tuberculosis H37Rv, M. bovis BCG, M. canetti, M. smegmatis, M. phlei, M. chelonae, M. kansasii, M. xenopi and M. avium were subjected to RD9 and 500 bp amplification by PCR. All M. tuberculosis strains, M. tuberculosis H37 Rv and M. canetti yielded a product of 333 bp which showed presence of RD9 region in these strains, whereas all M. bovis yielded a product of 206 bp with RD9 PCR assay. There was no ampli-fication product found in M. bovis BCG, M. xenopi, M. smegmatis, M. phlei, M. chelonae, M. kansasii, and M. avium. PCR based on 500 bp fragment showed a product of 500 bp in all M. bovis strains and M. bovis BCG. There was no amplification product of 500 bp found in M. canetti, M. smegmatis, M. phlei, M. chelonae, M. avium, M. kansasii, M. xenopi and was absent in all M. tuberculosis strains. The PCR assay results correlated 100% with the culture and biochemical results of the isolates. Our study suggested that PCR based on RD9 and 500 bp may effectively identify two closely related species of M. tuberculosis and M. bovis.
本研究通过RD9区域和500bp片段PCR检测对结核分枝杆菌和牛分枝杆菌进行鉴定。从40份人类痰液和41份牛肺标本中鉴定并表征了8株结核分枝杆菌和5株牛分枝杆菌，同时将保存在印度兽医研究所分枝杆菌实验室的20株结核分枝杆菌和9株牛分枝杆菌纳入本研究。通过这种方式，对28株结核分枝杆菌和14株牛分枝杆菌，以及作为比较和对照的结核分枝杆菌H37Rv、牛分枝杆菌卡介苗、堪氏分枝杆菌、耻垢分枝杆菌、草分枝杆菌、龟分枝杆菌、堪萨斯分枝杆菌、非洲分枝杆菌和鸟分枝杆菌进行了RD9和500bp的PCR扩增。所有结核分枝杆菌菌株、结核分枝杆菌H37Rv和堪氏分枝杆菌均产生了333bp的产物，表明这些菌株中存在RD9区域，而所有牛分枝杆菌通过RD9 PCR检测产生了206bp的产物。在牛分枝杆菌卡介苗、非洲分枝杆菌、耻垢分枝杆菌、草分枝杆菌、龟分枝杆菌、堪萨斯分枝杆菌和鸟分枝杆菌中未发现扩增产物。基于500bp片段的PCR在所有牛分枝杆菌菌株和牛分枝杆菌卡介苗中显示出500bp的产物。在堪氏分枝杆菌、耻垢分枝杆菌、草分枝杆菌、龟分枝杆菌、鸟分枝杆菌、堪萨斯分枝杆菌、非洲分枝杆菌中未发现500bp的扩增产物，且在所有结核分枝杆菌菌株中均不存在。PCR检测结果与分离株的培养和生化结果100%相关。我们的研究表明，基于RD9和500bp的PCR可有效鉴定结核分枝杆菌和牛分枝杆菌这两个密切相关的物种。
