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目的探讨TaqMan-MGB探针实时荧光PCR快速检测结核分枝杆菌北京基因型菌株的效果。方法对杭州地区136株结核分枝杆菌临床分离株水煮法提取DNA,分别用TaqMan-MGB探针实时荧光PCR与间隔区寡核苷酸(Spoligotyping)基因分型方法进行北京基因型菌株鉴定,比较2种方法的一致性,同时评价TaqMan-MGB探针实时荧光PCR的最低检测限。结果以Spoligotyping基因分型方法为标准,136株结核分枝杆菌中,检测到北京基因型结核分枝杆菌105株,占77.21%。TaqMan-MGB探针实时PCR方法与Spoligotyping基因分型方法鉴定北京基因型菌株结果完全一致。TaqMan-MGB探针实时荧光PCR对北京基因型最低检出限为0.488 pg/μl。结论与Spoligotyping基因分型方法相比,TaqMan-MGB探针实时PCR方法检测北京基因型操作简单,耗时短,灵敏度高,可以快速地区分北京基因型与非北京基因型结核分枝杆菌。
Objective To investigate the rapid detection of Mycobacterium tuberculosis Beijing genotype by TaqMan-MGB probe real-time PCR. Methods DNA was extracted from 136 clinical isolates of Mycobacterium tuberculosis in Hangzhou by boiling method. The genotypes of Beijing isolates were identified by TaqMan-MGB probe real-time PCR and Spoligotyping genotyping. The agreement of the two methods was compared, and the minimum detection limit of TaqMan-MGB probe real-time PCR was evaluated. Results According to the Spoligotyping genotyping method, 105 strains of Mycobacterium tuberculosis were detected in 136 strains of Mycobacterium tuberculosis in Beijing, accounting for 77.21%. TaqMan-MGB probe real-time PCR method and Spoligotyping genotyping method to identify the Beijing genotype strains exactly the same results. The detection limit of TaqMan-MGB probe real-time PCR in Beijing genotype was 0.488 pg / μl. Conclusion Compared with Spoligotyping, TaqMan-MGB real-time polymerase chain reaction (PCR) detection of Beijing genotypes is simple, time-consuming and sensitive. It can rapidly distinguish genotypes of Beijing from non-Beijing genotypes of Mycobacterium tuberculosis.