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

doi:10.19982/j.issn.1000-6621.20250031

Abstract

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

CLC Number:

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

Figures/Tables 4

References 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