High-resolution minisatellite-based typing as a portable approach to global analysis of Mycobacterium tuberculosis molecular epidemiology
High-resolution minisatellite-based typing as a portable approach to global analysis of Mycobacterium tuberculosis molecular epidemiology
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DOI：
10.1073/pnas.98.4.1901
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发表时间：
2001-02-13
影响因子：
11.1
通讯作者：
Supply, P
中科院分区：
文献类型：
--
作者：
Mazars, E;Lesjean, S;Supply, P
结核病对人类健康的全球性威胁强调了开发全球流行病学监测新方法的必要性。基于IS 6110限制性片段长度多态性（RFLP）的结核分枝杆菌分型的当前标准遭受在独立实验室之间比较数据的困难。在这里，我们提出了一个高分辨率的分型方法的基础上的可变数目串联重复序列（VNTR）的遗传元件命名为分枝杆菌散在重复单位（MIRU）在12个人类小卫星样区域的M。结核病基因组MIRU-VNTR图谱分析了72个不同的M.通过PCR分析所有12个位点建立结核分离株。在这些菌株的12个区域中鉴定出2至8个MIRU-VNTR等位基因，这对应于超过1600万种不同组合的潜力，产生接近IS 6110-RFLP的分辨率。MIRU-VNTR分型对所有流行病学相关分离株进行了完美聚类，表明这些MIRU-VNTR的稳定性足以跟踪暴发事件。MIRU-VNTR与IS 6110-RFLP分型结果之间的相关性极显著。与IS 6110-RFLP相比，MIRU-VNTR高分辨率分型方法具有快速、适用于所有M.结核病分离株，包括具有几个IS 6110拷贝的菌株，并允许容易和快速地比较来自独立实验室的结果。这种分型方法为结核分枝杆菌分子流行病学研究的数字化全球数据库的构建开辟了道路。
The worldwide threat of tuberculosis to human health emphasizes the need to develop novel approaches to a global epidemiological surveillance. The current standard for Mycobacterium tuberculosis typing based on IS6110 restriction fragment length polymorphism (RFLP) suffers from the difficulty of comparing data between independent laboratories. Here, we propose a high-resolution typing method based on variable number tandem repeats (VNTRs) of genetic elements named mycobacterial interspersed repetitive units (MIRUs) in 12 human minisatellite-like regions of the M. tuberculosis genome. MIRU-VNTR profiles of 72 different M. tuberculosis isolates were established by PCR analysis of all 12 loci. From 2 to 8 MIRU-VNTR alleles were identified in the 12 regions in these strains, which corresponds to a potential of over 16 million different combinations, yielding a resolution power close to that of IS6110-RFLP. All epidemiologically related isolates tested were perfectly clustered by MIRU-VNTR typing, indicating that the stability of these MIRU-VNTRs is adequate to track outbreak episodes. The correlation between genetic relationships inferred from MIRU-VNTR and IS6110-RFLP typing was highly significant. Compared with IS6110-RFLP, high-resolution MIRU-VNTR typing has the considerable advantages of being fast, appropriate for all M. tuberculosis isolates, including strains that have a few IS6110 copies, and permitting easy and rapid comparison of results from independent laboratories. This typing method opens the way to the construction of digital global databases for molecular epidemiology studies of M, tuberculosis.