新型PCR荧光探针技术用于结核病患者早期诊断的多中心应用评价研究

doi:10.19982/j.issn.1000-6621.20250031

中国防痨杂志 ›› 2025, Vol. 47 ›› Issue (6): 687-693.doi: 10.19982/j.issn.1000-6621.20250031

新型PCR荧光探针技术用于结核病患者早期诊断的多中心应用评价研究

欧喜超¹, 滕冲², 宋媛媛¹, 郑扬¹, 陈磊³, 朱俊⁴, 王建国⁵, 潘兆宝³, 康海涛⁴, 王彦⁵, 么鸿雁⁶, 黄飞¹()

¹传染病溯源预警与智能决策全国重点实验室,中国疾病预防控制中心结核病预防控制中心,北京 102206
²北京市东城区疾病预防控制中心结核科,公共卫生应急管理创新中心,北京 100050
³山东省潍坊市第二人民医院检验科,潍坊 261041
⁴河南省周口市传染病医院(周口市结核病防治所)检验科,周口 466001
⁵河北大学附属医院检验科,保定 071000
⁶中国疾病预防控制中心教育培训处,北京 102206

收稿日期:2025-01-20 出版日期:2025-06-10 发布日期:2025-06-11
通信作者: 黄飞,Email:huangfei@chinacdc.cn
作者简介:注:滕冲与欧喜超对本研究有同等贡献,为并列第一作者
基金资助:
国家重点研发计划(2022YFC2305204);国家重点研发计划(2023YFC2307301);西藏自治区重点研发计划(XZ202201ZY0007N-01);2528结核病预防控制项目

Multicenter evaluation study on the application of a novel PCR fluorescence probe technology for early diagnosis of tuberculosis

Ou Xichao¹, Teng Chong², Song Yuanyuan¹, Zheng Yang¹, Chen Lei³, Zhu Jun⁴, Wang Jianguo⁵, Pan Zhaobao³, Kang Haitao⁴, Wang Yan⁵, Yao Hongyan⁶, Huang Fei¹()

¹National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, National Center for Tuberculosis Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China
²Department of Tuberculosis, Beijing Dongcheng District Center for Disease Control and Prevention, Public Health Emergency Management Innovation Center, Beijing 100050, China
³Department of Laboratory Medicine, The Second People’s Hospital of Weifang, Shandong Province, Weifang 261041, China
⁴Clinical Laboratory, Zhoukou Communicable Disease Hospital (Zhoukou Tuberculosis Dispensary), Henan Province, Zhoukou 466001, China
⁵Clinical Laboratory, Affiliated Hospital of Hebei University, Baoding 071000, China
⁶Department of Education and Training, Chinese Center for Disease Control and Prevention, Beijing 102206, China

Received:2025-01-20 Online:2025-06-10 Published:2025-06-11
Contact: Huang Fei, Email: huangfei@chinacdc.cn
Supported by:
National Key Research and Development Program(2022YFC2305204);National Key Research and Development Program(2023YFC2307301);Key Research and Development Plan of Xizang Autonomous Region(XZ202201ZY0007N-01);2528 Tuberculosis Prevention and Control Project

摘要/Abstract

摘要：

目的: 评价新型PCR-荧光探针(DiagMed qPCR,简称“qPCR”)技术在初诊疑似肺结核患者中进行结核病诊断的可靠性,为其在结核病早期诊断领域的推广应用提供基础数据支持。方法: 选择潍坊市第二人民医院、周口市传染病医院和河北大学附属医院作为项目现场实施单位,采用前瞻性研究,连续纳入2024年6—10月期间到医院就诊且符合纳入标准的初诊疑似肺结核患者。实验室工作人员对纳入患者送检痰或支气管肺泡灌洗液样本进行涂片染色镜检、MGIT 960液体培养、GeneXpert MTB/RIF(简称“Xpert”)和qPCR检测。结果: 最终纳入563例初诊疑似肺结核患者,涂片镜检阳性率为38.01%(214/563),液体培养阳性率为50.80%(286/563),Xpert检测阳性率为54.53%(307/563),qPCR检测阳性率为58.79%(331/563),qPCR检测阳性率和Xpert检测阳性率均明显高于液体培养,差异均有统计学意义(χ²=22.000,P<0.001;χ²=9.468,P=0.003)。以液体培养结果为参照标准,qPCR检测结核分枝杆菌的敏感度和特异度分别为92.31%(264/286)和75.74%(206/272),Xpert检测结核分枝杆菌的敏感度和特异度分别为91.07%(255/280)和80.60%(216/268)。以临床诊断结果为参照标准,qPCR检测敏感度和特异度分别为68.05%(328/482)和96.30%(78/81),Xpert检测敏感度和特异度分别为65.04%(307/472)和100.00%(78/78)。结论: qPCR检测结核分枝杆菌具有非常好的检测效能,可用于在重点人群进行结核病患者主动发现,在综合医疗机构和结核病定点医疗机构进行结核病患者早期诊断。

关键词: 分枝杆菌,结核, 聚合酶链反应, 诊断,鉴别, 评价研究

Abstract:

Objective: To evaluate the reliability of the novel PCR fluorescence probe technique (DiagMed qPCR, abbreviated as “qPCR”) in diagnosing tuberculosis among tuberculosis suspected patients and provide foundational data support for its clinical application in early tuberculosis diagnosis. Methods: Prospective studies were conducted at The Second People’s Hospital of Weifang, Zhoukou Communicable Disease Hospital, and Affiliated Hospital of Hebei University. Patients meeting inclusion criteria who presented with suspected initial TB diagnosis between June and October 2024 were consecutively enrolled. Laboratory staff performed smear staining microscopy, MGIT 960 liquid culture, GeneXpert MTB/RIF (abbreviated as “Xpert”), and qPCR on sputum or bronchoalveolar lavage fluid (BALF) samples from enrolled patients. Results: A total of 563 patients with suspected initial TB diagnosis were included. The smear microscopy positivity rate was 38.01% (214/563), liquid culture positivity rate was 50.80% (286/563), Xpert positivity rate was 54.53% (307/563), and qPCR positivity rate was 58.79% (331/563). Both qPCR and Xpert positivity rates were significantly higher than that of liquid culture (χ²=22.000, P<0.001; χ²=9.468, P=0.003). Using liquid culture as the reference standard, qPCR demonstrated a sensitivity of 92.31% (264/286) and specificity of 75.74% (206/272), while Xpert showed a sensitivity of 91.07% (255/280) and specificity of 80.60% (216/268). Using clinical diagnosis as the reference standard, qPCR exhibited a sensitivity of 68.05% (328/482) and specificity of 96.30% (78/81), whereas Xpert demonstrated a sensitivity of 65.04% (307/472) and specificity of 100.00% (78/78). Conclusion: The qPCR technique demonstrates excellent diagnostic performance for Mycobacterium tuberculosis detection. It can be utilized for active case finding in key populations and for early TB diagnosis in general healthcare institutions and designated TB medical facilities.

Key words: Mycobacterium tuberculosis, Polymerase chain reaction, Diagnosis, differential, Evaluation studies

中图分类号:

欧喜超, 滕冲, 宋媛媛, 郑扬, 陈磊, 朱俊, 王建国, 潘兆宝, 康海涛, 王彦, 么鸿雁, 黄飞. 新型PCR荧光探针技术用于结核病患者早期诊断的多中心应用评价研究[J]. 中国防痨杂志, 2025, 47(6): 687-693. doi: 10.19982/j.issn.1000-6621.20250031

Ou Xichao, Teng Chong, Song Yuanyuan, Zheng Yang, Chen Lei, Zhu Jun, Wang Jianguo, Pan Zhaobao, Kang Haitao, Wang Yan, Yao Hongyan, Huang Fei. Multicenter evaluation study on the application of a novel PCR fluorescence probe technology for early diagnosis of tuberculosis[J]. Chinese Journal of Antituberculosis, 2025, 47(6): 687-693. doi: 10.19982/j.issn.1000-6621.20250031

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参考文献 15

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检测方法	液体培养(份)		合计 (份)	敏感度 (%,95%CI)	特异度 (%,95%CI)	阳性预测值 (%,95%CI)	阴性预测值 (%,95%CI)	Kappa值
检测方法	阳性	阴性	合计 (份)	敏感度 (%,95%CI)	特异度 (%,95%CI)	阳性预测值 (%,95%CI)	阴性预测值 (%,95%CI)	Kappa值
qPCR				92.31 (88.59~95.12)	75.74 (70.19~80.71)	80.00 (76.38~83.19)	90.35 (86.17~93.36)	0.674
阳性	264	66	330
阴性	22	206	228
合计	286	272	558
Xpert				91.07 (87.10~94.14)	80.60 (75.35~85.16)	83.06 (79.30~86.26)	89.63 (85.54~92.66)	0.718
阳性	255	52	307
阴性	25	216	241
合计	280	268	548
涂片镜检				66.20 (60.37~71.68)	90.44 (86.31~93.66)	87.85 (83.25~91.32)	71.93 (68.43~75.18)	0.563
阳性	188	26	214
阴性	96	246	342
合计	284	272	556

检测方法	液体培养(份)		合计 (份)	敏感度 (%,95%CI)	特异度 (%,95%CI)	阳性预测值 (%,95%CI)	阴性预测值 (%,95%CI)	Kappa值
检测方法	阳性	阴性	合计 (份)	敏感度 (%,95%CI)	特异度 (%,95%CI)	阳性预测值 (%,95%CI)	阴性预测值 (%,95%CI)	Kappa值
qPCR
痰液样本				93.55 (88.46~96.86)	64.00 (53.79~73.36)	80.11 (75.56~83.99)	86.49 (77.54~92.23)	0.603
阳性	145	36	181
阴性	10	64	74
合计	155	100	255
支气管肺泡灌洗液				90.84 (84.55~95.18)	82.56 (76.05~87.91)	79.87 (74.04~84.65)	92.21 (87.29~95.32)	0.722
阳性	119	30	149
阴性	12	142	154
合计	131	172	303
Xpert
痰液				93.96 (88.84~97.20)	71.88 (61.78~80.58)	83.33 (78.98~87.74)	88.46 (80.08~93.60)	0.681
阳性	140	27	167
阴性	9	69	78
合计	149	96	245
支气管肺泡灌洗液				87.79 (80.92~92.85)	85.47 (79.30~90.37)	82.14 (76.10~86.92)	90.18 (85.25~93.59)	0.727
阳性	115	25	140
阴性	16	147	163
合计	131	172	303

检测方法	临床诊断(例)		合计 (例)	敏感度 (%,95%CI)	特异度 (%,95%CI)	阳性预测值 (%,95%CI)	阴性预测值 (%,95%CI)
检测方法	结核	非结核	合计 (例)	敏感度 (%,95%CI)	特异度 (%,95%CI)	阳性预测值 (%,95%CI)	阴性预测值 (%,95%CI)
qPCR				68.05 (63.68~72.19)	96.30 (89.56~99.23)	99.09 (97.29~99.70)	33.62 (30.63~36.75)
阳性	328	3	331
阴性	154	78	232
合计	482	81	563
Xpert				65.04 (60.55~69.34)	100.00 (95.38~100.00)	100.00 (98.81~100.00)	32.10 (29.48~34.84)
阳性	307	0	307
阴性	165	78	243
合计	472	78	550

阳性级别	各组间平均秩次的差异值	Z值	调整后 P值
以“极低组”为参照
低组	61.27	3.630	0.001
中组	126.51	8.010	<0.001
高组	175.66	9.700	<0.001
以“低组”为参照
中组	65.24	5.280	<0.001
高组	114.38	7.520	<0.001
以“中组”为参照
高组	49.15	3.510	0.001

新型PCR荧光探针技术用于结核病患者早期诊断的多中心应用评价研究

Multicenter evaluation study on the application of a novel PCR fluorescence probe technology for early diagnosis of tuberculosis

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