Automated high-throughput genotyping for study of global epidemiology of Mycobacterium tuberculosis based on mycobacterial interspersed repetitive units
Automated high-throughput genotyping for study of global epidemiology of Mycobacterium tuberculosis based on mycobacterial interspersed repetitive units
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DOI：
10.1128/jcm.39.10.3563-3571.2001
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发表时间：
2001-10-01
影响因子：
9.4
通讯作者：
Locht, C
中科院分区：
文献类型：
--
作者：
Supply, P;Lesjean, S;Locht, C
大规模的结核分枝杆菌基因分型尤其具有挑战性，因为目前的分型方法是劳动密集型的，而且结果很难在不同的实验室之间进行比较。在这里，我们提出了12个哺乳动物小卫星样结核分枝杆菌基因座的可变数目串联重复序列(VNTRs)的自动分型方法。该系统将基于荧光的DNA分析仪上的多重PCR分析与基因分型的计算机自动化相结合。对来自38个国家的90个菌株的盲法参考集的分析(K.Kremer等人，J.Clin.微生物。37：2607-2618,1999)表明，该方法对结核分枝杆菌复杂分离株具有100%的重复性、敏感性和特异性，这是在相同条件下测试的任何其他分型方法所未达到的性能。MIRU-VNTRs可用于分析处于不同进化差异水平的结核分枝杆菌复合株的全球遗传多样性。为了充分利用该分型系统的可移植性，建立了一个通过互联网进行结核分枝杆菌Miru-VNTR基因分型的网站。这为结核病的全球流行病学监测开辟了道路，并应导致对这一主要病原体的进化和群体遗传学的新见解。
Large-scale genotyping of Mycobacterium tuberculosis is especially challenging, as the current typing methods are labor-intensive and the results are difficult to compare among laboratories. Here, automated typing based on variable-number tandem repeats (VNTRs) of genetic elements named mycobacterial interspersed repetitive units (MIRUs) in 12 mammalian minisatellite-like loci of M. tuberculosis is presented. This system combines analysis of multiplex PCRs on a fluorescence-based DNA analyzer with computerized automation of the genotyping. Analysis of a blinded reference set of 90 strains from 38 countries (K. Kremer et al., J. Clin. Microbiol. 37:2607-2618, 1999) demonstrated that it is 100% reproducible, sensitive, and specific for M. tuberculosis complex isolates, a performance that has not been achieved by any other typing method tested in the same conditions. MIRU-VNTRs can be used for analysis of the global genetic diversity of M. tuberculosis complex strains at different levels of evolutionary divergence. To fully exploit the portability of this typing system, a website was set up for the analysis of M. tuberculosis MIRU-VNTR genotypes via the Internet. This opens the way for global epidemiological surveillance of tuberculosis and should lead to novel insights into the evolutionary and population genetics of this major pathogen.