结核分枝杆菌耐药基因突变特征及与耐药水平关系的研究  

Study on characteristics of drug resistance gene mutation in Mycobacterium tuberculosisand its correlation with drug resistance level

作　　者：蒋燕成[1] 张建明 陈紫萱[1] 张志珊[1] JIANG Yancheng;ZHANG Jianming;CHEN Zixuan;ZHANG Zhishan(Department of Clinical Laboratory,Quanzhou First Affiliated Hospital of Fujian Medical University,Quanzhou,Fujian 362000,China)

机构地区：[1]福建医科大学附属泉州第一医院检验科,福建泉州362000

出　　处：《检验医学与临床》2023年第15期2145-2148,共4页Laboratory Medicine and Clinic

基　　金：福建省卫生健康委员会中青年骨干人才培养项目(2020GGA075);福建省泉州市科技计划项目(2018N066S)。

摘　　要：目的 分析抗结核一线药物利福平、异烟肼耐药基因rpoB、katG、ihnA的突变特征,以及基因突变位点与药物最低抑菌浓度(MIC)的相关性,以了解结核分枝杆菌(MTB)的耐药机制,指导临床用药。方法 收集泉州市第一医院2019年6月至2021年6月分离的耐利福平、异烟肼的MTB 51株作为研究对象,通过将菌种进行复苏,采用DNA微阵列芯片技术,快速检测MTB的耐药性。分析基因突变类型与抗结核药物MIC的关系。对药敏结果和基因芯片结果不相符合的标本进行测序,分析是否有新的耐药位点。结果 51株耐药MTB中有1株耐利福平的单耐药MTB,2株耐异烟肼的单耐药MTB,48株耐利福平、异烟肼的耐多药MTB。将48株MTB通过DNA微阵列芯片法进行基因型检测,发现rpoB中最常见的突变类型是531(C→T),异烟肼中最常见的是katG 315(G→C),其次是inhA-15(C→T)。将其中的23株MTB菌株进行MIC药敏试验,结果发现88.9%的531(C→T)突变菌株MIC>16μg/mL,5.6%的531(C→T)突变菌株MIC=16μg/mL,5.6%的531(C→T)突变菌株MIC=2μg/mL;4株526(C→T)突变型和1株516(A→T)突变型菌株的MIC均>16μg/mL。结论 泉州地区利福平最常见的突变类型是rpoB 531(C→T)突变型,异烟肼中最常见的是katG 315(G→C)突变型,rpoB密码子531、526、516突变类型对利福平产生高耐药水平;katG对异烟肼产生高耐药水平,inhA对异烟肼产生低耐药水平。Objective To analyze the mutation characteristics of rpoB,katG and ihnA of the first-line anti-tuberculosis drugs rifampicin and isoniazid resistance genes and gene mutation sites and drug MIC,so as to clarify the mechanism of drug resistance and guide medication.Methods Fifty-one strains of rifampicin-and isoniazid-resistant MTB were collected from June 2019 to June 2021 in Quanzhou First Hospital as the study objects,and MTB resistance was rapidly detected by resuscitating the strains and using DNA microarray chip technology.To analyze the relationship between gene mutation type and anti-tuberculosis drug minimal inhibitory concentration(MIC).Samples with inconsistent susceptibility and microarray results were sequenced to analyze for new resistance sites.Results Among the 51 strains of drug-resistant MTB,1 strain was of rifampicin-resistant MTB,2 strains were of isoniazid-resistant MTB,and 48 strains were of rifampicin-and isoniazid-resistant multidrug MTB.The genotype detection of 48 strains of MTB by DNA microarray method found that the most common mutation type in rpoB was 531(C→T),the most common of isoniazide was katG 315(G→C),followed by inhA-15(C→T).23 strains of MTB were subjected to MIC value susceptibility tests,the results showed that 88.9%of the 531(C→T)mutant strains had MIC>16μg/mL,5.6%had MIC=16μg/mL,5.6%had MIC=2μg/mL.1 strain of MTB with 526(C→T)mutant and 1 strain with 516(A→T)mutant strains had MIC values>16μg/mL.Conclusion The most common mutation type of rifampicin in Quanzhou area is rpoB 531(C→T)mutant,the most common type of isoniazid is katG 315(G→C)mutant,and rpoB codon 531,526 and 516 mutation types have high resistance levels to rifampicin.katG develops high resistance levels to isoniazid and inhA develop low levels of resistance to isoniazid.

关 键 词：结核分枝杆菌 耐药基因 最小抑菌浓度 

分 类 号：R52[医药卫生—内科学]