Value of DNA real-time fluorescence isothermal amplification method and combined testing in early diagnosis of tuberculous pleurisy

doi:10.19982/j.issn.1000-6621.20230055

Abstract

Abstract:

Objective: To investigate the value of DNA real-time fluorescence isothermal amplification method (abbreviated as “constant temperature amplification method”), combined with adenosine deaminase (ADA) and peripheral blood tuberculosis infection T cell spot test (T-SPOT. TB) detection in the early diagnosis of tuberculous pleurisy. Methods: Using retrospective research method, 427 patients with pleural effusion of unknown causes admitted to Xi’an Chest Hospital from January 2020 to December 2021 were selected as the research objects. All patients underwent pleural effusion isothermal amplification, ADA, Mycobacterium tuberculosis culture, and peripheral blood T-SPOT.TB testing. Based on clinical comprehensive diagnostic criteria as a reference, the early diagnostic value of pleural effusion DNA constant temperature amplification combined with ADA and peripheral blood T-SPOT.TB for tuberculous pleurisy was analyzed. Results: Among 427 subjects, 279 cases (65.3%) were diagnosed as tuberculous pleurisy (tuberculous pleurisy group) and 148 cases were diagnosed as non-tuberculous pleurisy (non-tuberculous pleurisy group). The optimal threshold of ADA was 19.5 U/L. Among the single detection methods, the specificity and sensitivity of the constant temperature amplification method were 100.00% and 31.54%, which were similar to those of Mycobacterium tuberculosis culture (100.00% and 30.11%, respectively). The sensitivity of ADA and T-SPOT.TB in peripheral blood was 74.91% and 84.95%,respectively, and the specificity was 87.16% and 74.91%, respectively. ADA+T-SPOT.TB could obtain high sensitivity (94.27%) and specificity (99.32%) in parallel and series, respectively. The parallel detection efficiency of isothermal amplification method, ADA and T-SPOT.TB got the same value as that with the Mycobacterium tuberculosis culture method in parallel. Among the joint detection methods, ADA+T-SPOT.TB detection in series had the highest cost-effectiveness, with a cost effectiveness ratio (C/E) of 669.23. Conclusion: The constant temperature amplification method with rapid detection protocols such as ADA and peripheral blood T-SPOT.TB could provide strong evidence for the early diagnosis and treatment of tuberculous pleurisy patients.

Key words: Mycobacterium tuberculosis, Pleurisy, Nucleic acid amplification techniques, Adenosine deaminase, Clinical laboratory techniques

CLC Number:

R521

Zhao Guolian, Lei Qian, Wang Pei, Ren Yunxia, Guan Jing. Value of DNA real-time fluorescence isothermal amplification method and combined testing in early diagnosis of tuberculous pleurisy[J]. Chinese Journal of Antituberculosis, 2023, 45(7): 681-686. doi: 10.19982/j.issn.1000-6621.20230055

Figures/Tables 5

References 21

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检测项目	结核性胸膜炎组 (279例)	非结核性胸膜炎组 (148例)	统计检验值	P值
恒温扩增法[例(构成比,%)]			χ²=58.799	<0.001
阳性	88(31.5)	0(0.0)
阴性	191(68.5)	148(100.0)
结核分枝杆菌培养[例(构成比,%)]			χ²=55.472	<0.001
阳性	84(30.1)	0(0.0)
阴性	195(69.9)	148(100.0)
胸腔积液ADA[U/L,M(Q₁,Q₃)]	37.0(19.0,48.0)	9.0(5.0,14.0)	U=6681.000	<0.001
外周血T-SPOT.TB检测[例(构成比,%)]			χ²=133.820	<0.001
阳性	237(84.6)	43(15.4)
阴性	42(25.4)	105(84.6)
胸腔积液LDH[U/L,M(Q₁,Q₃)]	396.0(230.0,598.0)	210.0(116.5,437.5)	U=13875.500	<0.001
外周血C反应蛋白[mg/L,M(Q₁,Q₃)]	14.7(6.9,35.2)	24.0(10.5,56.0)	U=16387.000	<0.001

指标	恒温扩增法	结核分枝杆菌培养	ADA检测	T-SPOT.TB 检测	联合方案1	联合方案2	联合方案3	联合方案4
成本C(元/次)	235	140	10	425	435	435	670	810
敏感度(%)	31.54	30.11	74.91	84.95	94.27	65.59	94.98	94.98
特异度(%)	100.00	100.00	87.16	70.95	58.78	99.32	58.78	58.78
效果E(约登指数)	0.32	0.30	0.62	0.56	0.53	0.65	0.54	0.54
C/E	734.38	466.67	16.13	758.93	820.75	669.23	1240.74	1500.00