Nanopore靶向测序在呼吸道样本中检测非结核分枝杆菌的诊断价值研究  

Diagnostic value of Nanopore targeted sequencing for detecting nontuberculous mycobacteria in respiratory specimens

作　　者：应广智 蔡青山[2] 马晓卿 陈凌燕[2] 陈园园[2] Ying Guangzhi;Cai Qingshan;Ma Xiaoqing;Chen Lingyan;Chen Yuanyuan(The Second School of Clinical Medicine,Zhejiang Chinese Medical University,Hangzhou 310053,China;Department of Tuberculosis,Hangzhou Red Cross Hospital,Hangzhou 310005,China)

机构地区：[1]浙江中医药大学第二临床医学院,杭州310053 [2]杭州市红十字会医院结核科,杭州310005

出　　处：《中国防痨杂志》2025年第5期589-596,共8页Chinese Journal of Antituberculosis

基　　金：国家自然科学基金(82104236)。

摘　　要：目的:探讨Nanopore靶向测序在呼吸道样本中检测非结核分枝杆菌(non-tuberculous mycobacteria,NTM)的诊断价值。方法:回顾性收集2021年12月至2022年9月在浙江省杭州市红十字会医院结核病诊疗中心行呼吸道样本Nanopore靶向测序的455例患者的资料。根据诊断标准,455例患者最终诊断为NTM-PD患者(NTM-PD组)65例,诊断为非NTM-PD患者(非NTM-PD组)390例,455例患者的肺泡灌洗液或痰液同时进行Nanopore靶向测序、BACTEC MGIT 960液体培养(简称“液体培养”)和(或)PCR荧光探针检测,比较各种方法诊断NTM-PD的效能。结果:以临床诊断结果为标准,Nanopore靶向测序、液体培养、PCR荧光探针检测呼吸道样本NTM的敏感度分别为92.3%(60/65)、83.1%(54/65)和57.1%(36/63),特异度分别为96.2%(375/390)、98.7%(385/390)和99.5%(380/382),阳性预测值分别为80.0%(60/75)、91.5%(54/59)和94.7%(36/38),阴性预测值分别为98.7%(375/380)、97.2%(385/396)和93.4%(380/407);Nanopore靶向测序、液体培养和PCR荧光探针的Kappa值分别为0.831、0.851和0.679。以液体培养为标准,Nanopore靶向测序和PCR荧光探针检测的敏感度分别为86.4%(51/59)和73.5%(36/49),特异度分别为98.0%(388/396)和94.4%(374/396),阳性预测值分别为86.4%(51/59)和62.1%(36/58),阴性预测值分别为98.0%(388/396)和96.6%(374/387);Nanopore靶向测序和PCR荧光探针的Kappa值分别为0.844和0.629。在65例临床诊断为NTM-PD组的患者中,Nanopore靶向测序和液体培养+基因芯片均为阳性的样本为50例,菌种鉴定的一致性为96.0%(48/50)。结论:Nanopore靶向测序在诊断NTM-PD中表现出较高的敏感度和特异度,与常规鉴定方法的一致性良好。在菌种鉴定方面,Nanopore靶向测序与液体培养结合基因芯片的检测结果具有高度一致性。Nanopore靶向测序能够对疑似NTM-PD患者的临床呼吸道样本进行快速检测,并获取菌种鉴定结果,为后续抗NTM治疗方案的制定提供指导。Objective:To investigate the diagnostic value of Nanopore targeted sequencing for detecting non-tuberculous mycobacteria(NTM)in respiratory samples.Methods:Data from 455 patients who underwent Nanopore targeted sequencing of respiratory samples from December 2021 to September 2022 at the Tuberculosis Treatment Center of Hangzhou Red Cross Hospital in Zhejiang Province was retrospectively collected.According to diagnostic criteria,these 455 patients were finally diagnosed as NTM-PD patients(NTM-PD group;65 patients)and non-NTM-PD patients(non-NTM-PD group;390 patients),their alveolar lavage fluid or sputum were subjected to simultaneous Nanopore targeted sequencing,BACTEC MGIT 960 liquid cultures(liquid culture)and/or PCR fluorescent probes to compare their efficacy in diagnosing NTM-PD.Results:Using clinical diagnostic results as standard,the sensitivities of Nanopore targeted sequencing,liquid culture,and PCR fluorescent probe for detecting NTM in respiratory samples were 92.3%(60/65),83.1%(54/65),and 57.1%(36/63),respectively;the specificities were 96.2%(375/390),98.7%(385/390),and 99.5%(380/382),respectively;and the positive predictive values were 80.0%(60/75),91.5%(54/59),and 94.7%(36/38);negative predictive values were 98.7%(375/380),97.2%(385/396),and 93.4%(380/407);Kappa values were 0.831,0.851 and 0.679,respectively.Using liquid culture as standard,the sensitivities of Nanopore targeted sequencing,and PCR fluorescent probe were 86.4%(51/59),and 73.5%(36/49),respectively;the specificities were 98.0%(388/396)and 94.4%(374/396),respectively;the positive predictive values were 86.4%(51/59)and 62.1%(36/58),respectively;and the negative predictive values were 98.0%(388/396)and 96.6%(374/387),respectively;Kappa values were 0.844 and 0.629,respectively.Among the 65 cases clinically diagnosed as NTM-PD,50 samples got positive results for both Nanopore targeted sequencing and liquid culture+gene chip,and their strain identification consistency was 96.0%(48/50).Conclusion:Nanopore targeted sequencing demonstrated hi

关 键 词：非典型性细菌 分枝杆菌感染 NANOPORES 诊断技术和方法 敏感性与特异性 

分 类 号：R446.5[医药卫生—诊断学]