靶向高通量测序在检测结核患者利福平和利福布汀耐药的应用研究  

Clinical application of targeted next generation sequencing in detecting rifampicin and rifabutin resistance in tuberculosis patients

作　　者：吴驰[1] 杨世瑾 赖文杰[1] 曲久鑫 Wu Chi;Yang Shijin;Lai Wenjie;Qu Jiuxin(Department of Clinical Laboratory,Shenzhen Third People′s Hospital,The Second Affiliated Hospital of Southern University of Science and Technology,National Clinical Research Center for Infectious Diseases,Shenzhen518112,China)

机构地区：[1]深圳市第三人民医院检验医学部,南方科技大学第二附属医院,国家感染性疾病临床医学研究中心,深圳518112

出　　处：《中华预防医学杂志》2024年第11期1673-1678,共6页Chinese Journal of Preventive Medicine

基　　金：广东省科技厅面上项目(2020B1111170014);深圳市科技计划项目(KCXFZ20230731100901003,KJZD20230923115116032)。

摘　　要：本研究探讨靶向高通量测序(tNGS)技术对诊断结核患者的利福平和利福布汀耐药的应用价值。采用回顾性队列研究,收集2020—2023年深圳市第三人民医院119例培养阳性结核患者的分枝杆菌菌株,tNGS检测rpoB基因耐药突变情况,其中46株结核分枝杆菌(MTB)中检出了14种不同型别的rpoB基因突变,包括43例rpoB基因耐药相关突变和3例529位密码子同义突变。以利福平及利福布汀最小抑菌浓度(MIC)表型药敏结果为金标准,tNGS方法检测利福平和利福布汀耐药的敏感度均为100%,特异度为96.2%和89.4%,具有良好的诊断性能。其中rpoB基因531、526位突变与利福平和利福布汀的耐药高度相关。在此基础上,tNGS检测23例结核患者临床原始痰标本与对应菌株结果一致,对18例多次分枝杆菌培养阳性患者的83株MTB菌株进行tNGS检测,结果显示2例MTB初始无突变患者后续检出rpoB基因突变同时利福平和利福布汀表型药敏变为耐药。综上,tNGS检测可显示rpoB基因突变类型,减少同义突变造成的假阳性结果,对诊断利福平和利福布汀耐药具有良好的检验性能;tNGS可直接检测临床痰标本,还可用于动态监测MTB分子耐药谱变化。To evaluate the clinical value of targeted next generation sequencing(tNGS)in diagnosing rifampicin and rifabutin resistance in tuberculosis patients.In this retrospective cohort study,119 culture-positive Mycobacterium tuberculosis(MTB)strains from tuberculosis patients in Shenzhen Third People′s Hospital from 2020 to 2023 were collected,then tNGS was performed to detect mutations of rpoB gene.Fourteen different types of rpoB gene mutation were detected in 46 mutation MTB strains,including 43 resistance related mutations and 3 synonymous mutations at codon 529.Using the phenotypic drug susceptibility results of rifampicin and rifabutin as the reference standard,the sensitivities of tNGS for detecting resistance to rifampicin and rifabutin were 100%,and the specificities were 96.2%and 89.4%respectively,therefore,tNGS showed good diagnostic performance.Mutations at positions 531 and 526 of rpoB were highly associated with resistance to rifampicin and rifabutin.Moreover,the results of tNGS from the clinical specimens were consistent with those from the corresponding culture strains.tNGS analysis was performed on 83 MTB strains from 18 patients with multiple positive cultures.The results showed that 2 patients with no mutations in the initial MTB strains were subsequently detected with rpoB gene mutation and their phenotypic drug susceptibilities changed from sensitive to resistant.In summary,using tNGS to detect rpoB mutations can reduce false positive results caused by synonymous mutations,and have satisfactory performance for the diagnosis of rifampicin and rifabutin resistance.tNGS can directly detect clinical sputum samples,and also can be used to dynamically monitor the molecular resistance profiles of MTB,therefore it has extremely broad clinical application prospects.

关 键 词：结核分枝杆菌 靶向高通量测序 利福平 耐药 

分 类 号：R52[医药卫生—内科学]