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目的分析青岛地区结核分枝杆菌embB306基因突变规律,探讨embB306基因突变与耐多药的相关性。方法根据结核分枝杆菌embB基因设计扩增引物和测序引物,PCR扩增embB基因,利用焦磷酸测序技术测定89株结核分枝杆菌临床分离株embB306基因位点序列,采用Spearman相关分析法分析embB306基因位点的突变与耐多药的相关性。结果 23株耐多药结核菌株(MDR-TB)中,14株发现embB306基因突变,突变比例为60.9%;46株非MDRTB中,7株发现embB306基因突变,突变比例为15.2%,突变率在MDR-TB组与非MDR-TB组差异有统计学意义(χ~2=15.09,P﹤0.01),Spearman相关分析显示结核分枝杆菌embB306基因突变与耐多药呈正相关(r=0.471,P﹤0.01),主要突变形式为ATG→GTG和ATG→ATC。结论青岛地区MDR-TB菌株的embB306基因突变率显著高于非MDRTB菌株,embB306突变与耐多药具有相关性。
Objective To analyze the mutation rule of embB306 gene in Qingdao and to explore the relationship between embB306 gene mutation and multidrug resistance. Methods According to the embB gene of M. tuberculosis, the primers for embB gene were designed and the embB gene was amplified by PCR. The sequence of embB306 gene of 89 clinical isolates of Mycobacterium tuberculosis was detected by pyrosequencing technique. The expression of embB306 gene was analyzed by Spearman correlation analysis Mutations in gene loci correlate with multidrug resistance. Results Among the 23 MDR-TB strains, 14 strains of embB306 gene mutation were found in 60 strains of MDR-TB strains, of which embB306 gene mutation was found in 7 of 46 non-MDRTB strains, the mutation rate was 15.2%. The mutation rate was The difference between MDR-TB group and non-MDR-TB group was statistically significant (χ ~ 2 = 15.09, P <0.01). Spearman correlation analysis showed that the mutation of embB306 in M. tuberculosis was positively correlated with MDR (r = 0.471, P <0.01). The main mutations were ATG → GTG and ATG → ATC. Conclusion The mutation rate of embB306 gene in MDR-TB strains in Qingdao is significantly higher than that in non-MDRTB strains, and embB306 mutation is associated with multidrug resistance.