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目的研究结核分枝杆菌(Mycobacterium tuberculosis,MTB)emb B306位点及其他突变位点与乙胺丁醇(ethambutol,EMB)的耐药表型及耐多药(multidrug resistant,MDR)的关系;分析emb B基因突变与EMB药敏表型及MDR的关系。方法对临床分离的MTB采用BD MGIT 960 SIRE试剂比例法进行药敏试验,取48株EMB耐药、46株EMB敏感但耐其他药及7株四药均敏感MTB提取核酸并扩增emb B基因全序列,对扩增产物进行测序分析emb B基因序列,与H37Rv标准株序列比对,分析emb B基因各突变位点、形式及频率。结果 101株MTB发现emb B基因序列上有17个不同位点突变形式。53株MTB在emb B基因序列上发生突变,其中46株为EMB耐药,7株为EMB敏感;emb B基因野生型的MTB有48株,其中2株为EMB耐药,46株为EMB敏感;emb B突变型与emb B野生型的MTB之间EMB耐药率有显著性差异(c2=68.95,P<0.01)。101株MTB中MDR有51株,其中有42株发生emb B突变,9株为emb B基因野生型,emb B基因突变率在MDR和非MDR之间有显著性差异(c2=36.9,P<0.01)。结论 emb B306位点与EMB耐药及MDR中度相关,emb B基因突变与EMB耐药及耐多药结核菌(multidrug resistant-Mycobacterium tuberculosis,MDR-TB)高度相关,emb B基因突变可作为MDR-TB的检测分子标记物,指导临床用药。
Objective To study the relationship between embB306 and other mutation sites in Mycobacterium tuberculosis (MTB) and the resistant phenotype of ethambutol (EMB) and multidrug resistant (MDR) The relationship between emb B mutation and EMB susceptibility phenotype and MDR. Methods MTB assay was used to detect the susceptibility of MTB with BD MGIT 960 SIRE reagent. 48 EMB-resistant, 46 EMB-sensitive but resistant to other drugs and 7 strains of four-drug sensitive MTB were used to extract nucleic acid and amplify the emb B gene The sequence of the emb B gene was sequenced and compared with the H37Rv standard strain. The site, the form and the frequency of the emb B gene mutation were analyzed. Results 101 strains of MTB found emb B gene sequence of 17 different sites on the mutant form. Fifty-three strains of MTB mutated in the emb B gene sequence, of which 46 were EMB-resistant and 7 were EMB-sensitive; 48 strains of embB gene wild-type MTB, of which 2 were EMB-resistant and 46 were EMB-sensitive There was a significant difference in EMB resistance between emb B mutant and emb B wild-type MTB (c2 = 68.95, P <0.01). There were 51 MDR strains in 101 MTBs, of which 42 were emb B mutations and 9 were wild-type emb B genes. The mutation rate of emb B gene was significantly different between MDR and non-MDR (c2 = 36.9, P < 0.01). Conclusion The emb B306 locus is moderately associated with EMB resistance and MDR. The emb B gene mutation is highly correlated with EMB resistance and multidrug resistant Mycobacterium tuberculosis (MDR-TB). The emb B gene mutation may be used as MDR -TB detection of molecular markers, to guide clinical medication.