北京基因型耐多药结核分枝杆菌二线抗结核药物耐药基因突变特征分析  

Second-line drug resistance associated mutations in multidrug-resistant Mycobacterium tuberculosis Beijing genotype strains

作　　者：叶静芬 方晴 胡耀仁 许小敏 车洋[2] YE Jingfen;FANG Qing;HU Yaoren;XU Xiaomin;CHE Yang(Department of Infection,Hwa Mei Hospital,University of Chinese Academy of Sciences,Ningbo,Zhejiang 315010,China;不详)

机构地区：[1]中国科学院大学宁波华美医院感染科,浙江宁波315010 [2]宁波市疾病预防控制中心结核病防制所

出　　处：《预防医学》2021年第10期983-987,共5页CHINA PREVENTIVE MEDICINE JOURNAL

基　　金：宁波市省市共建医学重点学科传染病学(2016-S04)。

摘　　要：目的了解耐多药结核分枝杆菌(MDR-TB)北京基因型菌株对二线抗结核药物的耐药情况及耐药基因突变特征。方法对中国科学院大学宁波华美医院2017—2019年临床分离的MDR-TB采用RD105缺失基因检测法鉴定基因型。采用药敏试验比例法检测MDR-TB对卡那霉素、阿米卡星、卷曲霉素、氧氟沙星和左氧氟沙星等二线抗结核药物的耐药性。以结核分枝杆菌标准株H37RV为参比序列,PCR法检测rrs、tlyA、eis、gidB、gyrA和gyrB耐药基因突变,通过比较不同基因型的耐药率和基因突变率,分析北京基因型二线抗结核药物耐药特征。结果133株MDR-TB经鉴定,北京基因型106株,非北京基因型27株。北京基因型MDR-TB卡那霉素、阿米卡星、卷曲霉素、氧氟沙星和左氧氟沙星耐药率分别为9.43%、7.55%、3.77%、32.08%和32.08%,准广泛耐药率为30.19%,广泛耐药率为7.55%,与非北京基因型比较差异均无统计学意义(P>0.05)。北京基因型MDR-TB rrs、tlyA、eis、gidB、gyrA和gyrB突变率分别为7.55%、7.55%、1.89%、2.83%、36.79%和2.83%,与非北京基因型比较差异均无统计学意义(P>0.05)。rrs突变类型有2种,tlyA 1种,eis 2种,gidB 2种,gyrA 5种,gyrB 3种,均以点突变为主。结论北京基因型菌株为MDR-TB的优势菌株,对氟喹诺酮类药物耐药率较高,以gyrA基因突变为主。Objective To learn the characteristics of second-line drug resistance and related gene mutations of multidrug-resistant Mycobacterium tuberculosis(MDR-TB)Beijing genotype strains.Methods The MDR-TB isolates in Hwa Mei Hospital from 2017 to 2019 were enrolled and detected using RD105 deletion-targeted multiplex polymerase chain reaction(PCR).The proportion method for drug susceptibility test was used to detect the drug-resistant profiles against kanamycin,amikacin,capreomycin,ofloxacin and levofloxacin.The gene sequencing of rrs,tlyA,eis,gidB,gyrA and gyrB was conducted by PCR compared with H37RV strain.The differences in the rates of drug resistance and mutation between Beijing and non-Beijing genotype strains were examined to understand the characteristics of Beijing genotype strains.Results There were 106 Beijing genotype and 27 non-Beijing genotype strains in 133 MDR-TB isolates.The drug resistance rates of kanamycin,amikacin,capreomycin,ofloxacin and levofloxacin in Beijing genotype strains were 9.43%,7.55%,3.77%,32.08%and 32.08%,respectively.The rates of quasi-extensive and extensive drug resistance in Beijing genotype strains were 30.19%and 7.55%.The gene mutation rates of rrs,tlyA,eis,gidB,gyrA and gyrB in Beijing genotype strains were 7.55%,7.55%,1.89%,2.83%,36.79%and 2.83%,respectively.There were no significantly differences between Beijing and Non-Beijing genotype strains in the factors above(P>0.05).The gene rrs,tlyA,eis,gidB,gyrA and gyrB had 2,1,2,2,5 and 3 mutation types,respectively,with single base substitution as the main type.Conclusion Beijing genotype strains are dominant in MDR-TB,with high resistance to fluoroquinolones and mainly gyrA gene mutation.

关 键 词：结核分枝杆菌 北京基因型 耐药 基因突变 

分 类 号：R521[医药卫生—内科学]