纳米孔测序技术对非结核分枝杆菌肺病的诊断价值研究

doi:10.19983/j.issn.2096-8493.20250024

结核与肺部疾病杂志 ›› 2025, Vol. 6 ›› Issue (4): 413-419.doi: 10.19983/j.issn.2096-8493.20250024

纳米孔测序技术对非结核分枝杆菌肺病的诊断价值研究

黄高燕, 苏深小, 赵娟娟, 梁宝元, 蒙夏艳(), 何华伟

南宁市第四人民医院/广西艾滋病临床治疗中心(南宁)结核科,南宁 530021

收稿日期:2025-01-23 出版日期:2025-08-20 发布日期:2025-08-08
通信作者: 蒙夏艳,Email:2540089309@qq.com
基金资助:
广西壮族自治区卫生健康委员会自筹经费科研课题(Z-A20231204)

Study on the diagnostic value of nanopore sequencing technology for non-tuberculous mycobacterial pulmonary disease

Huang Gaoyan, Su Shenxiao, Zhao Juanjuan, Liang Baoyuan, Meng Xiayan(), He Huawei

Department of Tuberculosis, Nanning Fourth People’s Hospital/Guangxi Clinical Treatment Center for AIDS (Nanning), Nanning 530021,China

Received:2025-01-23 Online:2025-08-20 Published:2025-08-08
Contact: Meng Xiayan, Email: 2540089309@qq.com
Supported by:
Self-funded Research Project of Guangxi Zhuang Autonomous Region Health Commission(Z-A20231204)

摘要/Abstract

摘要：

目的: 探讨纳米孔测序技术对疑似非结核分枝杆菌(non-tuberculous mycobacterial,NTM)肺病患者支气管肺泡灌洗液的诊断价值。方法: 选取2022年1月1日至2024年12月31日期间,在南宁市第四人民医院结核科就诊的疑似NTM肺病患者155例,经支气管镜检查收集支气管肺泡灌洗液(bronchoalveolar lavage fluid,BALF)进行分枝杆菌培养及鉴定、分枝杆菌鉴定(荧光PCR熔解曲线法)及纳米孔测序技术检测,以最终确诊NTM肺病为标准比较三种结果的诊断效率。结果: 最终确诊NTM肺病患者96例(NTM肺病组),非NTM肺病组患者59例(诊断肺结核29例、其他肺炎30例),纳米孔测序对NTM肺病诊断的敏感度、诊断一致率、Kappa值分别为92.71%(95%CI:85.98%~96.75%)、86.45%(95%CI:79.81%~91.56%)、0.706(95%CI:0.586~0.808),明显高于分枝杆菌鉴定(荧光PCR熔解曲线法)[63.54%(95%CI:53.18%~73.09%)、71.61%(95%CI:63.91%~78.49%)、0.445(95%CI:0.350~0.618)]、分枝杆菌培养及鉴定[63.54%(95%CI:53.18%~73.09%)、74.84%(95%CI:67.35%~81.44%)、0.515(95%CI:0.453~0.703)],以及后两者联合检测[84.38%(95%CI:75.53%~90.94%)、82.58%(95%CI:75.67%~88.23%)、0.634(95%CI:0.523~0.761)];纳米孔测序法的特异度[76.27%(95%CI:63.60%~86.37%)]低于分枝杆菌鉴定(荧光PCR熔解曲线法)[84.75%(95%CI:72.90%~92.71%)]、分枝杆菌培养及鉴定[93.22%(95%CI:83.54%~98.01%)、后两者联合检测79.66%(95%CI:67.34%~89.14%)]。结论: 纳米孔测序技术对NTM肺病具有较高的诊断效率,能够早期快速检出非结核分枝杆菌,并对菌种的鉴定有优势,有诊断NTM肺病的应用价值,但在临床应用中应考虑特异度问题。

关键词: 纳米孔测序, 非典型性细菌, 分枝杆菌属, 支气管肺泡灌洗液

Abstract:

Objective: To explore the diagnostic value of nanopore sequencing technology for detecting non-tuberculous mycobacterial pulmonary disease (NTM-PD) in bronchoalveolar lavage fluid (BALF). Methods: A total of 155 patients with suspected NTM-PD admitted to the Tuberculosis Department of Nanning Fourth People’s Hospital from January 1, 2022, to December 31, 2024, were enrolled. BALF samples collected via bronchoscopy were subjected to mycobacterial culture and identification, fluorescence PCR melting curve method, and nanopore sequencing. The diagnostic efficiencies of these three methods were compared. Results: Among the patients, 96 were diagnosed with NTM-PD (NTM-PD group) and 59 with non-NTM pulmonary diseases (including 29 cases of pulmonary tuberculosis and 30 cases of other pneumonias). The sensitivity, diagnostic concordance rate, and Kappa value of nanopore sequencing for NTM-PD diagnosis were 92.71% (95%CI: 85.98%-96.75%), 86.45% (95%CI: 79.81%-91.56%), and 0.706 (95%CI: 0.586-0.808),The sensitivity of nanopore sequencing was significantly higher than that of mycobacterium identification by fluorescence PCR melting curve method (63.54%,95%CI:53.18%-73.09%;71.61%,95%CI:63.91%-78.49%;0.445,95%CI:0.350-0.618), mycobacterium culture and identification (63.54%,95%CI:53.18%-73.09%;74.84%,95%CI:67.35%-81.44%;0.515,95%CI:0.453-0.703), and the combination of the latter two methods (84.38%,95%CI:75.53%-90.94%;82.58%,95%CI:75.67%-88.23%;0.634,95%CI:0.523-0.761). However,the specificity of nanopore sequencing was 76.27%(95%CI:63.60%-86.37%), which was lower than that of mycobacterium identification by fluorescence PCR melting curve method (84.75%,95%CI:72.90%-92.71%), mycobacterium culture and identification (93.22%,95%CI:83.54%-98.01%), and the combination of the latter two methods (79.66%,95%CI:67.34%-89.14%). Conclusion: Nanopore sequencing technology has high diagnostic efficiency for NTM-PD and can directly identify bacterial species, but specificity issues should be considered in clinical application.

Key words: Nanopore sequencing, Atypical bacteria forms, Mycobacterium, Bronchoalveolar lavage fluid

中图分类号:

R517.9

黄高燕, 苏深小, 赵娟娟, 梁宝元, 蒙夏艳, 何华伟. 纳米孔测序技术对非结核分枝杆菌肺病的诊断价值研究[J]. 结核与肺部疾病杂志, 2025, 6(4): 413-419. doi: 10.19983/j.issn.2096-8493.20250024

Huang Gaoyan, Su Shenxiao, Zhao Juanjuan, Liang Baoyuan, Meng Xiayan, He Huawei. Study on the diagnostic value of nanopore sequencing technology for non-tuberculous mycobacterial pulmonary disease[J]. Journal of Tuberculosis and Lung Disease, 2025, 6(4): 413-419. doi: 10.19983/j.issn.2096-8493.20250024

图/表 3

表1

两组患者一般情况

特征	NTM肺病组(96例)	非NTM肺病组(59例)	统计检验值	P值
性别[例,构成比(%)]			χ²=1.310^a	0.253
男性	35(36.46)	27(45.76)
女性	61(63.54)	32(54.24)
年龄(岁, $x ˉ$ ±s)	59.37±12.04	58.82±12.95	t=-0.268	0.789
吸烟史[例,率(%)]	6(6.25)	2(3.39)	χ²=0.607^a	0.436
既往病史[例,率(%)]
高血压	9(9.38)	7(11.86)	χ²=0.243^a	0.622
糖尿病	4(4.17)	4(6.78)	χ²=0.506^a	0.477
慢性肾病	10(10.42)	6(10.17)	χ²=0.0137^a	0.907
艾滋病	2(2.08)	1(1.69)	χ²=0.0289^a	0.865
支气管扩张症	58(60.42)	19(32.20)	χ²=11.560^a	0.001

表1

表2

表3

参考文献 14

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特征	例数	率(%)	特征	例数	率(%)
临床症状			腰痛	2	2.08
咳嗽	64	66.67	气喘	5	5.21
咳痰	55	57.29	头痛、头晕	2	2.08
胸闷、气促	14	14.58	呕吐	1	1.04
咯血	10	10.42	皮肤疱疹	1	1.04
发热	5	5.21	颈淋巴结肿大并破溃感染	1	1.04
纳差	4	4.17	影像学征象
乏力	5	5.21	支气管扩张	58	60.42
消瘦	3	3.13	空洞影	23	23.96
胸痛	4	4.17	多发结节	47	48.96

纳米孔测序技术对非结核分枝杆菌肺病的诊断价值研究

Study on the diagnostic value of nanopore sequencing technology for non-tuberculous mycobacterial pulmonary disease

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检测方法	NTM肺病诊断		敏感度 (%)	特异度 (%)	阳性预测值(%)	阴性预测值(%)	一致率 (%)	Kappa值
检测方法	阳性(例)	阴性(例)	敏感度 (%)	特异度 (%)	阳性预测值(%)	阴性预测值(%)	一致率 (%)	Kappa值
纳米孔测序技术			92.71	76.27	86.41	86.54	86.45	0.706
阳性	89	14
阴性	7	45
分枝杆菌鉴定(荧光PCR熔解曲线法)			63.54	84.75	87.14	58.82	71.61	0.445
阳性	61	9
阴性	35	50
分枝杆菌培养及鉴定			63.54	93.22	93.85	61.11	74.84	0.515
阳性	61	4
阴性	35	55
联合检测方法^a			84.38	79.66	87.10	75.81	82.58	0.634
阳性	81	12
阴性	15	47

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[5]	张志学苍爱泽孙静. 应用GeneXpert MTB/RIF技术检测肺泡灌洗液对菌阴肺结核的临床诊断价值[J]. 结核病与肺部健康杂志, 2017, 6(2): 128-130.
[6]	唐怡敏,叶涛生,邓国防,等. 艾滋病并发菌阴肺结核诊治中支气管肺泡灌洗液检测的应用进展[J]. 结核病与肺部健康杂志, 2017, 6(1): 68-72.