熔解曲线法检测结核分枝杆菌耐药在唐山地区的应用  

Application of melting curve method to detect drug resistance of Mycobacterium tuberculosis in Tangshan Area

作　　者：刘海涛 田杰 赵俊鹏[2] 韩云艳 周志枫 张燕 LIU Haitao;TIAN Jie;ZHAO Junpeng;HAN Yunyan;ZHOU Zhifeng;ZHANG Yan(Department of Clinical Laboratory Tangshan Seventh Hospital,Tangshan,Hebei 063000,China;不详)

机构地区：[1]唐山市第七医院检验科,河北唐山063000 [2]唐山市疾病预防控制中心结核科

出　　处：《医学动物防制》2024年第5期459-462,共4页Journal of Medical Pest Control

基　　金：河北省2022年度医学科学研究重点课题计划(20221806)。

摘　　要：目的评价熔解曲线法检测结核分枝杆菌(Mycobacterium tuberculosis,MTB)对利福平和异烟肼耐药的效能,分析唐山地区MTB的耐药特征,为临床治疗结核病提供依据。方法通过表型药敏(微孔板法)、分子药敏(熔解曲线法)分别对275株MTB利福平和异烟肼的耐药性进行检测,经Kappa检验进行两组结果的比对分析。结果唐山地区利福平和异烟肼的表型耐药率分别为14.18%、28.64%;分子耐药率分别为17.95%、26.01%。以微孔板法为参考方法,熔解曲线法检测利福平耐药的敏感性、特异性、符合率分别为89.74%、94.02%、93.41%,与微孔板法一致性良好(Kappa=0.754);异烟肼耐药的敏感性、特异性、符合率分别为76.32%、93.40%、88.64%,与微孔板法一致性中等(Kappa=0.656)。利福平和异烟肼耐药基因突变存在多样性,利福平耐药基因突变以rpoB基因529~533位密码子单区域突变为主,突变比例53.06%(26/49);异烟肼耐药基因突以katG315密码子突变为主,突变比例53.52%(38/71)。结论熔解曲线法可快速、准确地检测MTB对多种药物的耐药性,可作为药敏检测的可靠方法推广应用。Objective To evaluate the efficacy of the melting curve method in detecting the resistance of Mycobacterium tuberculosis(MTB)to rifampicin and isoniazid,and to analyze the drug resistance characteristics of MTB in Tangshan Area,to provide a basis for clinical treatment of tuberculosis.Methods The resistance of 275 MTB strains of rifampicin and isoniazid was detected by phenotypic antimicrobial susceptibility(microplate method)and molecular antimicrobial susceptibility(melting curve method),respectively,and the results of the two groups were compared and analyzed by the Kappa test.Results The phenotypic resistance rates of rifampicin and isoniazid in Tangshan Area were 14.18%and 28.64%.The rates of molecular resistance were 17.95%and 26.01%.Using the microplate method as the reference method,the sensitivity,specificity,compliance of the melting curve method for the detection of rifampicin resistance were 89.74%,94.02%,and 93.41%,respectively,which were consistent with the microplate method(Kappa=0.754).The sensitivity,specificity,and coincidence rates of isoniazid resistance were 76.32%,93.40%,and 88.64%,respectively,which were of moderate consistency with the microplate method(Kappa=0.656).There was a diversity of rifampicin and isoniazid resistance gene mutations,and the rifampicin resistance gene mutations were mainly codon single region mutations at positions 529-533 of the rpoB gene,with a mutation ratio of 53.06%(26/49).The codon mutation of katG315 was the main mutation of the isoniazid resistance gene,and the mutation proportion was 53.52%(38/71).Conclusion The melting curve method can quickly and accurately detect the resistance of MTB to a variety of drugs,and can be popularized and applied as a reliable method for drug susceptibility detection.

关 键 词：结核分枝杆菌 耐药结核 分子耐药 熔解曲线法 PCR 

分 类 号：R446.5[医药卫生—诊断学]