IS6110-RFLP、Spoligotyping和MLVA在结核分枝杆菌基因分型中的应用研究

目的 评价156110限制性片段长度多态性（IS6110-RFLP）、间隔区寡核苷酸分型（Spoligotyping）及多位点可变数量串联重复序列分析（MLVA）3种方法在结核分枝杆菌基因分型研究中的应用。方法 分别采用IS6110-RFLP、Spoligotyping及MLVA对40株结核分枝杆菌临床分离株进行基因分型研究，比较3种方法的分型效果。结果 使用IS6110-RFLP方法，40株结核分枝杆菌呈现出35种基因型，33株具有独特的基因型；使用Spoligotyping方法，40株结核分枝杆菌呈现出17种基因型，11株具有独特的基因型；使用MLVA方法时，40株结核分枝杆菌呈现出34种基因型，29株具有独特的基因型；当3种方法联合使用时，40株结核分枝杆菌呈现出39种基因型，38株具有独特的基因型。结论 MLVA和IS6110-RFLP方法的分辨率高于Spoligotyping，但是对于IS6110单拷贝或低拷贝的菌株，MLVA方法的分辨率要高于IS6110-RFLP。MLVA、IS6110-RFLP和Spoligotyping联合应用可有效地进行结核病的流行病学调查和病原学监测。

The application of IS6110-RFLP, Spoligotyping and MLVA analysis in genotyping of Mycobacterium tuberculosis strains

Objective To evaluate the application of IS6110-restriction fragment length polymorphism (RFLP),spacer oligotyping(Spoligotyping)and multiple locus VNTR analysis in epidemiological studies of tuber- culosis.Methods Forty clinical isolates of M.tuberculosis were typed by IS6110-RFLP,Spoligotyping and VN- TR respectivdy to compare the results of the three methods.Results The result of IS6110-RFLP showed that 40 strains of M.tuberculosis present 35 of genotypes,33 strains have unique genotypes.The result of Spoligotyping showed that 40 strains present 17 of genotypes,11 strains have unique genotypes.The result of VNTR showed that 40 strains present 34 kinds of genotypes,29 strains have unique genotypes.The combination of IS6110-RFLP, Spoligotyping and VNTR showed that 40 strains present 39 kinds of genotypes,38 strains have unique genotypes. Conclusion That combined IS6110-RFLP Spoligotyping and MLVA are very useful for the epidemiological investi- gation of tuberculosis and its pathological surveillance.