间隔区寡核苷酸分型和多位点可变数量串联重复序列分析在结核分枝杆菌基因分型中的应用

目的评价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)方法在结核分枝杆菌基因分型研究中的应用。方法收集224株结核分枝杆菌临床分离株,分别采用Spoligotyping及MLVA方法进行基因分型,比较两种方法的分型效果,评价两种方法在结核分枝杆菌基因分型中的应用。结果使用Spoligotyping方法,224株结核分枝杆菌呈现出55种基因型,39株具有独特的基因型,其余185株菌呈现出16种基因型;使用MLVA方法时,224株结核分枝杆菌呈现出160种基因型,132株具有独特的基因型,余下的92株菌呈现出28种基因型;当两种方法联合使用时,224株结核分枝杆菌呈现出179种基因型,159株结核分枝杆菌具有独特的基因型,余下的65株菌表现为20种基因型。湖南省和安徽省的菌株中北京家族菌株所占的比例差异有统计学意义(P<0.001),安徽省北京家族菌株所占的比例明显高于湖南省。结论MLVA在结核分枝杆菌株水平的鉴定方面,其分辨能力高于Spoligotyping,但是Spoligotyping在鉴定北京家族菌株和M.bovis方面有一定的优势。将Spoligotyping方法作为一线分型技术,MLVA作为二线分型技术联合应用时,将提高结核病的流行病学调查和病原学监测效果。不同地区的菌株有不同的特点。

Application of Spoligotyping and MLVA analysis in genotype studies of Mycobacterium tuberculosis

Objective To access the application of spacer oligotyping(Spoligotyping) and Multiple Locus VNTR(MLVA) in epidemiological studies of Mycobacterium tuberculosis.Methods 224 clinical isolates of M.tuberculosis were collected and typed by Spoligotyping and MLVA respectively,to compare the results of both methods and to access their application in epidemiological studies of M.tuberculosis. Results Data from Spoligotyping showed that 224 strains presented 55 kinds of genotypes.Of these,39 were represented by a unique isolate,with the remaining 185 isolates being grouped in 16 clusters whereas the result of MLVA showed that 224 strains presenting 160 kinds of genotypes.Of these,132 were represented by a unique isolate,with the remaining 92 isolates being grouped in 28 groups.Data from the combination of Spoligotyping and VNTR showed that 224 strains presenting 179 kinds of genotypes.Of these,159 were represented by a unique isolate,with the remaining 65 isolates being grouped in 20 groups. There was significant difference noticed among M.tuberculosis between Hunan and Anhui in the proportion of Beijing family(P<0.001).The proportion of Beijing family in Anhui was higher than that in Hunan. Conclusion Results from this direct comparison studies demonstrated that MLVA analysis was more effective than Spoligotyping in discriminating individual M.tuberculosis isolates.However,Spoligotyping had an advantage over MLVA in identifying Beijing family strains and M.bovis.Taking Spoligotyping as a first-line typing technique and VNTR as second-line typing technique,the arrangement would improve the effectiveness of epidemiological investigation and pathological inspection of tuberculosis.The strains in different regions seemed to have had different characteristics.