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目的初步评价间隔区寡核苷酸分型(Spoligotyping)技术在鉴定结核分枝杆菌多重感染中的应用。方法选取结核分枝杆菌临床分离株,5μm孔径滤膜过滤法制备单个细菌菌悬液,平板接种37℃培养,选取生长良好的单个菌落培养增菌,提取基因组DNA,Spoligotyping进行基因分型鉴定,比较临床分离株与相应的单克隆分离株的分型结果。结果共选取32株结核分枝杆菌临床分离株,获得306个单克隆分离株。Spoligotyping分型鉴定结果显示30株结核分枝杆菌临床分离株与相应的单克隆分离株的分型结果一致,2株临床分离株的单克隆分离株呈现了多种基因型,多重感染率为6.25%。结论 Spoligotyping可用于快速,有效地区分结核分枝杆菌多重感染。
Objective To evaluate the application of Spoligotyping in the identification of multiple Mycobacterium tuberculosis infection. Methods A single bacterial suspension was prepared by clinical isolates of Mycobacterium tuberculosis and filtered by 5μm pore size filter. The single bacterial suspension was inoculated into the plate at 37 ℃. Single well-growing colonies were selected and cultured for enrichment. Genomic DNA and Spoligotyping were extracted for genotyping. The typing results of the clinical isolates and the corresponding monoclonal isolates were compared. Results A total of 32 clinical isolates of Mycobacterium tuberculosis were selected and 306 monoclonal isolates were obtained. The results of Spoligotyping typing showed that the clinical isolates of 30 strains of Mycobacterium tuberculosis were consistent with the typing results of the corresponding monoclonal isolates. The monoclonal isolates of 2 clinical isolates showed multiple genotypes and the multiple infection rate was 6.25 %. Conclusion Spoligotyping can be used to quickly and effectively distinguish multiple infections with M. tuberculosis.