青岛地区结核分枝杆菌embB306基因突变与耐多药相关性研究

Study on the association between embB codon 306 mutation and multidrug resistance tuberculosis in Qingdao

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 characteristics of embB codon 306 mutation of Mycobacterium tuberculosis（MTB） in Qingdao by pyrosequencing technology and to explore the association between embB codon 306 mutation and MDR-TB. Methods To design the amplifying primer and sequencing primer according to M. tuberculosis embB gene. The fragments of embB306 gene of all of the 89 M. tuberculosis isolates were amplified,embB306 gene was detected by pyrosequencing in 89 strains. The association between embB306 mutation and MDR-TB in Qingdao were statistically analyzed by Spearman correlation analysis. Results 60.9%（14/23） of MDR-TB strains and 15.2%（7/46）of non-resistant strains had mutations at embB306. The embB306 mutation rate in multidrug resistant strains is sig-nificantly higher than that from non-multidrug resistant strains（χ~2=15.09,P 〈0.01）. Spearman correlation analysis showed that embB306 mutation had significant correlation with MDR-TB（r=0.471,P 〈0.01）. The mutation of ATG→GTG and ATG→ATC were the common mutant type. Conclusion There was a strong association between the embB306 mutation and MDR-TB in Qingdao.