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首页> 外文期刊>Journal of Clinical Microbiology >Automated High-Throughput Genotyping for Study of Global Epidemiology of Mycobacterium tuberculosis Based on Mycobacterial Interspersed Repetitive Units
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Automated High-Throughput Genotyping for Study of Global Epidemiology of Mycobacterium tuberculosis Based on Mycobacterial Interspersed Repetitive Units

机译：基于分枝杆菌重复单元的自动化高通量基因分型研究结核分枝杆菌的全球流行病学

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摘要

Large-scale genotyping of Mycobacterium tuberculosisis 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 ofM. 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 ofM. 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.
机译：结核分枝杆菌的大规模基因分型尤其具有挑战性，因为当前的分型方法需要大量劳动，而且很难在实验室之间进行比较。在此，基于12个哺乳动物M型迷你卫星样基因座中名为分枝杆菌散布重复单元（MIRU）的遗传元件的可变数目串联重复序列（VNTR）的自动分型。介绍了结核病。该系统将基于荧光的DNA分析仪上多重PCR的分析与基因分型的计算机自动化结合在一起。对来自38个国家/地区的90个菌株的盲参考集的分析（K. Kremer等人，J。Clin。Microbiol。37：2607–2618，1999）表明，它对具有100％的重现性，敏感性和特异性。 M.结核病复杂分离株，在相同条件下测试过的任何其他分型方法均未达到该性能。 MIRU-VNTR可用于分析 M的全球遗传多样性。肺结核复杂菌株在不同水平的进化分歧。为了充分利用此键入系统的可移植性，建立了一个网站来分析 M。互联网上的结核 MIRU-VNTR基因型。这为结核病的全球流行病学监测开辟了道路，并应导致对该主要病原体的进化和种群遗传学的新颖见解。

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