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目的对临床分离的耐多药结核分枝杆菌株进行基因突变分析。方法对耐多药结核分枝杆菌临床分离菌株进行耐药基因的PCR检测,利用基因序列测定方法分析耐药基因突变情况。结果从耐多药结核病(MDR-TB)患者临床分离菌株中快速检测到rpoB、katG、rpsL、embB基因及其突变。耐多药菌株、全耐及单耐利福平分离菌株均检测rpoB基因点突变,突变位点主要为第516、526和531常见密码子,1例MDR-TB出现第479位和第531位密码子同时突变。耐多药菌、全耐菌及单耐异烟肼的菌株均检测有katG基因点突变,突变位点均为第2066位碱基C突变为G。全耐菌和对乙胺丁醇(EMB)耐药的耐多药菌株均检测有embB基因点突变,突变位点均为第306位密码子ATG突变为ACG。全耐菌和对链霉素(SM)耐药的MDR-TB菌株均检测有rpsL基因点突变,突变密码子为CCT突变为CTT,其中从1株对SM敏感的MDR-TB中也检测到突变。全敏感株、标准株及利福平(RIF)、异烟肼(INH)或EMB敏感的耐药株均无rpoB、katG或embB基因突变。结论PCR及基因序列测定可快速检测耐多药结核基因,结核分枝杆菌耐多药性与多个基因突变相关。
Objective To analyze the gene mutation of multidrug-resistant Mycobacterium tuberculosis strains isolated clinically. Methods The clinical isolates of multidrug-resistant Mycobacterium tuberculosis were detected by polymerase chain reaction (PCR) and the gene mutations were analyzed by gene sequencing. Results Rapid detection of rpoB, katG, rpsL and embB genes and their mutations in clinical isolates from patients with multidrug-resistant tuberculosis (MDR-TB). Multidrug resistant strains, resistant and resistant to Rifampicin isolates were detected point mutations in rpoB gene mutations mainly 516,526 and 531 common codons, 1 case of MDR-TB appeared 479 and 531 Codon mutation at the same time. The multidrug-resistant, full-resistant and single-isoniazid-resistant isolates all detected katG gene point mutation, and the mutation site was the 2066th base C mutation. Mutations in the embB gene were detected in the full-resistant and multi-drug resistant strains to ethambutol (EMB). The site of mutation was the ATG mutation at codon 306 of ACG. Mutations in the rpsL gene were detected in fully resistant and susceptible streptomycin (SM) -treated MDR-TB strains, with the mutation codon CCT mutated to CTT, of which MDR-TB was also detected in a SM-sensitive SM mutation. There were no mutations of rpoB, katG or embB in all sensitive strains, standard strains and rifampin (RIF), isoniazid (INH) or EMB-sensitive resistant strains. Conclusion PCR and gene sequencing can detect MDR-TB gene rapidly. The multi-drug resistance of Mycobacterium tuberculosis is related to multiple gene mutations.