Sajduda Anna, Dziadek Jarosław, Kotłowski Roman, Portaels Françoise
Department of Genetics of Microorganisms, University of Łódz, Łódz 90-237, Poland.
Diagn Microbiol Infect Dis. 2006 May;55(1):59-64. doi: 10.1016/j.diagmicrobio.2005.12.004. Epub 2006 Feb 20.
In the present study, 77 drug-resistant Mycobacterium tuberculosis strains isolated in Poland in 2000 were characterized by the mycobacterial interspersed repetitive unit-variable number tandem repeat (MIRU-VNTR) typing and our novel method based on PCR amplification of DNA regions between IS6110 and 16-bp GC-rich frequent repeats (designated IS6110-Mtb1/Mtb2 PCR). The results were compared with previous data of the more commonly used methods, IS6110 restriction fragment length polymorphism (RFLP) and spoligotyping. The discriminatory power of IS6110-Mtb1/Mtb2 method was only slightly lower than that of IS6110 RFLP, whereas MIRU-VNTR typing was the least discriminative among the 4 methods used. Clustering of strains by using results of IS6110-Mtb1/Mtb2 PCR correlated well with RFLP-defined clusters, further confirming epidemiologic relationships among patients. These results indicate that the novel genotyping method could be an attractive alternative for other PCR-based typing procedures, such as spoligotyping and MIRU-VNTR typing. Also, it seems to be a valuable adjunct to the reference IS6110 RFLP method for studying the genetic diversity of drug-resistant M. tuberculosis strains in Poland.
在本研究中，对2000年在波兰分离出的77株耐多药结核分枝杆菌菌株进行了分枝杆菌散布重复单位可变数目串联重复序列（MIRU-VNTR）分型，以及基于对IS6110和富含GC的16 bp频繁重复序列之间的DNA区域进行PCR扩增的新方法（命名为IS6110-Mtb1/Mtb2 PCR）。将结果与更常用方法IS6110限制性片段长度多态性（RFLP）和间隔寡核苷酸分型法（spoligotyping）的先前数据进行了比较。IS6110-Mtb1/Mtb2方法的鉴别力仅略低于IS6110 RFLP，而MIRU-VNTR分型是所使用的4种方法中鉴别力最低的。利用IS6110-Mtb1/Mtb2 PCR结果对菌株进行聚类与RFLP定义的聚类相关性良好，进一步证实了患者之间的流行病学关系。这些结果表明，这种新的基因分型方法可能是其他基于PCR的分型方法（如间隔寡核苷酸分型法和MIRU-VNTR分型法）的有吸引力的替代方法。此外，对于研究波兰耐多药结核分枝杆菌菌株的遗传多样性，它似乎是参考IS6110 RFLP方法的有价值的辅助方法。
