Medical University of Lodz, Lodz, Poland.
Int J Tuberc Lung Dis. 2011 Sep;15(9):1252-8, i. doi: 10.5588/ijtld.10.0718.
Mycobacterium tuberculosis is one of the most dangerous human pathogens. Molecular typing of M. tuberculosis has allowed better control of tuberculosis and, among other benefits, identification of genetic lineages among strains.
To test the potential of polymerase chain reaction (PCR) based methods for the epidemiological study of M. tuberculosis strains isolated from patients residing in a single city.
We performed spoligotyping, mycobacterial interspersed repetitive units-variable number of tandem repeats (MIRU-VNTR) typing and insertion sequence (IS) 6110 restriction fragment length polymorphism (RFLP) analyses of 234 clinical strains of M. tuberculosis collected over 2 years from the Polish city of Lodz.
Spoligotyping analysis revealed 84 spoligotypes with a shared international type and 50 unique spoligotypes. Subtyping via 15- and 19-loci MIRU-VNTR analyses revealed 154 patterns with 117 unique profiles, and 159 patterns with 126 unique profiles, respectively. Spoligotyping combined with MIRU-VNTR 15- and 19 loci analyses revealed 132 and 146 unique profiles, respectively. Overall, 96 strains clustered via MIRU-VNTR typing were used in IS6110-RFLP analysis. Complete congruence of patterns revealed by PCR-based methods was noted for 40 strains, of which 36 were isolated from epidemiologically linked patients.
The combination of 15-loci MIRU-VNTR typing with spoligotyping is useful for primary analysis of M. tuberculosis strains; however, additional use of MIRU 23 should be considered. Strains clustered by PCR-based methods should be further analysed by IS6110-RFLP typing.
结核分枝杆菌是最危险的人类病原体之一。结核分枝杆菌的分子分型有助于更好地控制结核病，并且除了其他好处之外，还可以鉴定菌株之间的遗传谱系。
测试基于聚合酶链反应（PCR）的方法在对来自单个城市的患者分离的结核分枝杆菌菌株进行流行病学研究中的潜力。
我们对 2 年来从波兰罗兹市采集的 234 株临床结核分枝杆菌菌株进行 spoligotyping、分枝杆菌插入序列 6110 （IS6110） 限制片段长度多态性（RFLP）分析和 15 个和 19 个基因座的微卫星重复可变数串联重复（MIRU-VNTR）分型。
spoligotyping 分析显示 84 种 spoligotype 具有共同的国际型，50 种独特 spoligotype。通过 15 个和 19 个基因座 MIRU-VNTR 分析进行的亚分型显示，分别有 154 种和 146 种具有 117 种和 126 种独特图谱。 spoligotyping 与 15 个和 19 个基因座 MIRU-VNTR 分析相结合分别显示了 132 种和 146 种独特图谱。总体而言，使用 MIRU-VNTR 分型聚类的 96 株菌用于 IS6110-RFLP 分析。基于 PCR 的方法显示的模式完全一致的菌株为 40 株，其中 36 株从具有流行病学关联的患者中分离。
15 个基因座 MIRU-VNTR 分型与 spoligotyping 相结合可用于结核分枝杆菌菌株的初步分析；但是，应考虑进一步使用 MIRU23。基于 PCR 的方法聚类的菌株应进一步通过 IS6110-RFLP 分型进行分析。
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