纳米孔测序技术在耐药结核病快速诊断中的应用价值OA
Application value of nanopore sequencing technology in the rapid diagnosis of drug-resistant tuberculosis
目的 验证纳米孔测序技术在检测临床标本中结核分枝杆菌(Mycobacterium tuberculosis,M.tb)耐药性的价值.方法 收集2023年2月至2024年1月长春市传染病医院就诊的136例疑似耐药结核病患者的临床样本(痰、支气管肺泡灌洗液、胸水等)进行纳米孔测序检测,并将纳米孔测序检测结果与MGIT960液体培养、快速分子检测(Xpert MTB/RIF)结果进行比对.以常规表型药敏试验结果为参考标准,分析纳米孔测序法对结核分枝杆菌耐药性的快速诊断价值.结果 纳米孔测序法发现利福平耐药基因突变主要在rpoB S531L位点上,突变率为68.57%(72/105);异烟肼耐药基因突变主要在KatG S315T、inhA c15t位点上,突变率分别为56.41%(44/78)、26.92%(21/78);乙胺丁醇耐药基因突变主要在embB M306V、embB M306I位点上,突变率分别为40.00%(20/50)、24.00%(12/50);链霉素耐药基因突变主要在rpsL K43R位点上,突变率为77.14%(54/70);喹诺酮类药物耐药基因突变主要在gyrA D94G、gyrA A90V位点上,突变率分别为36.06%(22/61)、27.86%(17/61).纳米孔测序技术检测利福平、异烟肼、乙胺丁醇、链霉素、喹 诺 酮 类 的 敏 感 度 分 别 为:100%、92.21%、92.45%、87.14%、91.53%,特异度分别是为:83.78%、88.14%、98.80%、86.36%、90.91%,准确率分别是:95.59%、90.44%、96.32%、86.76%、91.18%,Kappa值分别为0.88、0.80、0.92、0.74、0.82.结论 纳米孔测序技术在耐药结核病快速检测中有较高的应用价值,能够满足临床及时诊断耐药结核病的需求.
This study was aimed at verifying the value of nanopore sequencing technology in detecting the drug resistance of My-cobacterium tuberculosis(M.tb)in clinical specimens from Changchun Infectious Disease Hospital.We retrospectively collected 136 clinical samples(e.g.,sputum,bronchoalveolar lavage fluid,or pleural effusion)from patients with suspected drug-resistant tubercu-losis between February 2023 and January 2024 at our hospital.Specimens were analyzed with nanopore sequencing.The results were compared with those for MGIT960 liquid culture and Xpert MTB/RIF,and conventional phenotypic drug susceptibility testing served as the reference standard.The rifampicin resistance mutations occurred primarily at rpoB S531L(68.57%,72/105);isoniazid resis-tance mutations occurred primarily at KatG S315T(56.41%,44/78)and inhAc15t(26.92%,21/78);ethambutol resistance mutations occurred primarily at embB M306V(40.00%,20/50)and embB M306I(24.00%,12/50);streptomycin resistance mutations occurred primarily at rpsLK43R(77.14%,54/70);and quinolone resistance mutations occurred primarily at gyrA D94G(36.06%,22/61)and gyrA A90V(27.86%,17/61).For these respective drugs,nanopore technology demonstrated a sensitivity of 100%,92.21%,92.45%,87.14%,and 91.53%;specificity of 83.78%,88.14%,98.80%,86.36%,and 90.91%;accuracy of 95.59%,90.44%,96.32%,86.76%,and 91.18%;and Kappa values of 0.88,0.80,0.92,0.74,and 0.82 in detecting resistance.Our results confirmed that nano-pore sequencing technology has high application value for rapid detection of drug-resistant tuberculosis and can meet the clinical needs for timely diagnosis at medical institutions.
李杨;孙晓柯;张健;曹阳;段鸿飞
首都医科大学附属北京胸科医院,北京 101149||长春市传染病医院,长春 130123河南省胸科医院结核内科,郑州 450008长春市传染病医院,长春 130123长春市传染病医院,长春 130123首都医科大学附属北京胸科医院,北京 101149
医药卫生
耐药结核病诊断方法快速检测纳米孔测序点突变评价研究
drug-resistant tuberculosisdiagnostic methodsrapid detectionnanopore sequencingpoint mutationevalua-tion study
《中国人兽共患病学报》 2026 (4)
362-368,7
北京市高层次公共卫生技术人才建设项目(No.G2024-2-003)河南省科技厅科技发展计划(No.232102310306) Programme for the Development of High-level Public Health Technical Talent in Beijing(No.G2024-2-003)Science and Technology Development Plan of the Science and Technology Department of Henan Province(No.232102310306)
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