靶向捕获测序技术和宏基因组二代测序技术检测肺泡灌洗液中结核分枝杆菌的诊断价值  

Diagnostic value of probe capture-based targeted next-generation sequencing and metagenomic next-generation sequencing for detecting Mycobacterium tuberculosis in bronchoalveolar lavage fluid

作　　者：石玉如 谷德健 吴静[1] 刘婷 秦令寒 岳莉[1] 戚应杰[1] Shi Yuru;Gu Dejian;Wu Jing;Liu Ting;Qin Linghan;Yue Li;Qi Yingjie(Anhui Provincial Hospital Infection District(Hefei Infectious Disease Hospital),Hefei 230000,China;Beijing Jiyinjia Technology Co.,Ltd,Beijing 102206,China)

机构地区：[1]安徽省立医院感染病院(合肥市传染病医院),合肥230000 [2]北京吉因加科技有限公司,北京102206

出　　处：《中国防痨杂志》2025年第3期305-311,共7页Chinese Journal of Antituberculosis

基　　金：合肥市应用医学项目(Hwk2023yb010)。

摘　　要：目的:探讨靶向捕获测序技术和宏基因组二代测序技术在疑似肺结核患者肺泡灌洗液标本中,对结核分枝杆菌的诊断价值。方法:回顾性分析2023年6月至2024年3月间,以安徽省立医院感染病院结核病院区收治的117例疑似肺结核患者的肺泡灌洗液作为研究样本,对其进行宏基因组二代测序、靶向捕获测序、GeneXpert MTB/RIF(以下简称“Xpert”)、涂片及培养,分析结核分枝杆菌检出情况,评价宏基因组测序和靶向测序两种方法对肺结核的检测效能。结果:117名疑似肺结核患者中,最终确诊为肺结核的患者为71例(60.68%),非肺结核患者46例(39.32%)。采用宏基因组二代测序、靶向捕获测序、涂片+培养、涂片+培养+Xpert四种检测方法的敏感度分别为77.46%(55/71)、76.06%(54/71)、34.29%(24/70)、54.93%(39/71),特异度分别为100.00%(46/46)、95.65%(44/46)、95.65%(44/46)、95.65%(44/46),阳性预测值分别为100.00%(55/55)、96.43%(54/56)、92.31%(24/26)、95.12%(39/41),阴性预测值分别为74.19%(46/62)、72.13%(44/61)、48.89%(44/90)、57.89%(44/76),符合率分别为86.32%(101/117)、83.76%(98/117)、58.62%(68/116)、70.94%(83/117),约登指数分别为0.77、0.72、0.30、0.51。宏基因组二代测序与靶向捕获测序检出结核分枝杆菌RPM值分别为0.22(0.02,1.95)和28.54(1.65,138.16),差异有统计学意义(Z=6.338,P<0.05)。靶向捕获测序检测利福平、氟喹诺酮、异烟肼耐药基因结果与表型耐药的符合率为100.00%。结论:宏基因组二代测序技术与靶向捕获测序技术在肺结核诊断中较传统检测方法都有着较好的诊断效能;同时靶向捕获测序技术还可以进行耐药位点检测,为临床用药提供参考。Objective:To explore the diagnostic value of probe capture-based targeted next-generation sequencing(tNGS)and metagenomic next-generation sequencing(mNGS)in bronchoalveolar lavage fluid for detection of pulmonary tuberculosis.Methods:A retrospective study was conducted from the bronchoalveolar lavage fluid of 117 presumed pulmonary tuberculosis patients admitted to the Tuberculosis Department of Anhui Provincial Hospital during June 2023 to March 2024 was retrospectively analyzed.The patients received tNGS,mNGS,acid fast bacilli smear microscopy,culture and Xpert MTB/RIF.The performance of tNGS and mNGS for detection of Mycobacterium tuberculosis was evaluated.Results:Among 117 suspected tuberculosis patients,71 patients(60.68%)were diagnosed with pulmonary tuberculosis,while 46 cases(39.32%)were non tuberculosis patients.The sensitivity of mNGS,tNGS,smear combined with culture,smear combined with culture and Xpert MTB/RIF were 77.46%(55/71),76.06%(54/71),34.29%(24/70)and 54.93%(39/71)respectively.The specificity were 100.00%(46/46),95.65%(44/46),95.65%(44/46)and 95.65%(44/46).The positive predictive values were 100.00%(55/55),96.43%(54/56),92.31%(24/26)and 95.12%(39/41),while the negative predictive values were 74.19%(46/62),72.13%(44/61),48.89%(44/90)and 57.89%(44/76).The accordance rate were 86.32%(101/117),83.76%(98/117),58.62%(68/116)and 70.94%(83/117),and the Youden index were 0.77,0.72,0.30 and 0.51.The performance of the first two groups was higher than that of the last two groups.The RPM values of Mycobacterium tuberculosis detected by mNGS and tNGS were 0.22(0.02,1.95)and 28.54(1.65,138.16),respectively,with statistically significant difference(Z=6.338,P<0.05).The accordance rate of detection of resistance to rifampicin,fluoroquinolone,and isoniazid between tNGS and phenotypic method was 100.00%.Conclusion:mNGS and tNGS have better diagnostic efficacy in the diagnosis of pulmonary tuberculosis compared to traditional detection methods.At the same time,tNGS can also be used for drug resistance detection,

关 键 词：结核 肺 肺泡灌洗液 分子诊断技术 诊断 鉴别 

分 类 号：R521[医药卫生—内科学] R49[医药卫生—临床医学]