三种不同的DNA分型技术在158株结核杆菌研究中的应用

目的评价IS6110限制性片段长度多态性(RFLP)、间隔区寡核苷酸分型(Spoligotyping)及分枝杆菌散在重复单位(MIRU)三种分型方法在结核病流行病学研究中的应用。方法对158株结核分枝杆菌临床分离株应用IS6110RFLP、Spoligotyping及MIRU三种分型方法进行鉴定。结果应用三种分型方法产生的类型数分别为118、20和105个。IS6110RFLP的分辨率大于Spoligotyping,MIRU的分辨能力与IS6110RFLP接近。在MIRU的12个区中,重复区4、10、26、40具有较高的多态性。广东地区与其他地区成簇率和北京基因型所占比例差异有统计学意义(P<0.05),广东地区成簇率和北京基因型所占比例均显著低于其他地区。结论应用IS6110RFLP、Spoligotyping及MIRU三种分型方法进行结核病流行病学研究具有重要意义且非常有效,可以发现中国不同地区菌株的不同特点。

The identification of Mycobacterium tuberculosis isolates by DNA typing technique

Objective To assess the application of IS6110-restriction fragment length polymorphism (RFLP), Spoligotyping and mycobacterial interspersed repetitive unit(MIRU) in epidemiological studies of tuberculosis and to discuss the characteristics of Mycobacterium tuberculosis strains in different regions in China. Methods Mycobacterium tuberculosis strains, with a total number of 158 isolates, were subjected to IS6110-RFLP, Spoligotyping and MIRU. Results The numbers of patterns produced by IS6110-RFLP, Spoligotyping, and MIRU typing were 118, 20 and 105 respectively. The discriminatory power of IS6110-RFLP was higher than that of Spoligotyping. However, when the copies of IS6110 were lower than 10, the discriminatory power of Spoligotyping improved obviously. The discriminatory power of MIRU typing was close to that of IS6110-RFLP for typing of Mycobacterium tuberculosis . In MIRU loci, there were four loci(loci 4,10,26,40) with higher diversity. Significant differences among the Mycobacterium tuberculosis between Guangdong and other regions in clustered rate and the proportion of Beijing genotype ( P < 0.05 ) were found. The clustered rates and the proportion of Beijing genotype in Guangdong were lower than that in other regions. Conclusion The results of this study indicated that either IS6110-RFLP, Spoligotyping or MIRU technique was useful for epidemiological studies on tuberculosis in China and the strains in different regions had different characterishes in China.