Allele-specific PCR method based on pncA and oxyR sequences for distinguishing Mycobacterium bovis from Mycobacterium tuberculosis:: Intraspecific M-bovis pncA sequence polymorphism
Allele-specific PCR method based on pncA and oxyR sequences for distinguishing Mycobacterium bovis from Mycobacterium tuberculosis:: Intraspecific M-bovis pncA sequence polymorphism
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DOI：
10.1128/jcm.36.1.239-242.1998
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发表时间：
1998-01-01
影响因子：
9.4
通讯作者：
Blázquez, J
中科院分区：
文献类型：
--
作者：
de los Monteros, LEE;Galán, JC;Blázquez, J
本文研究了一种基于两种基因多态性的等位基因特异性扩增方法，用于区分结核分枝杆菌和牛分枝杆菌。基于在来自M.结核和M. bovis的PCR系统，该系统能够区分237个M.在两个物种中的一个中测试的结核病复合群分离物的开发。所有121 M。结核菌株在169位显示预期的碱基（胞嘧啶）。大部分的M。牛分离物在所述位置具有鸟嘌呤。然而，116个M中的18个。牛结核分枝杆菌分离株（均为山羊结核分枝杆菌分离株）均表现出结核分枝杆菌特有的pncA多态性。这些结果表明，山羊牛分枝杆菌可能是分枝杆菌起源的烟酰胺酶缺失环节。结核菌种类基于oxyR基因285位的多态性，用同样的系统来区分M。结核分枝杆菌M.牛在这种情况下，DNA从所有121 M。结核分离株在该位置具有预期的碱基（鸟嘌呤）。此外，所有116 M。牛的分离株，包括那些家山羊，显示出相同的多态性（腺嘌呤）。oxyR等位基因特异性扩增法可区分M. bovis从M.结核病，是快速的（结果可以在不到3小时内获得），并易于执行。
An allele-specific amplification method based on two genetic polymorphisms to differentiate Mycobacterium tuberculosis from Mycobacterium bovis was tested. Based on the differences found at position 169 in the pncA genes from M. tuberculosis and M. bovis, a PCR system which was able to differentiate most of the 237 M. tuberculosis complex isolates tested in one of the two species was developed. All 121 M. tuberculosis strains showed the expected base (cytosine) at position 169. Most of the M. bovis isolates had a guanine at the cited position. Nevertheless, 18 of the 116 M. bovis isolates, all of them goat isolates, showed the pncA polymorphism specific to M tuberculosis. These results suggest that goat M bovis may be the nicotinamidase-missing link at the origin of the M. tuberculosis species. Based on the polymorphism found at position 285 in the oxyR gene, the same system was used to differentiate M. tuberculosis from M. bovis. In this case, DNAs from all 121 M. tuberculosis isolates had the expected base (guanine) at this position. In addition, all 116 M. bovis isolates, including those hom goats, showed the identical polymorphism (adenine). The oxyR allele-specific amplification method can differentiate M. bovis from M. tuberculosis, is rapid (results can be obtained in less than 3 h), and is easy to perform.