DNA实时荧光恒温扩增法联合检测对结核性胸膜炎的早期诊断价值

doi:10.19982/j.issn.1000-6621.20230055

摘要/Abstract

摘要：

目的: 分析DNA实时荧光恒温扩增法(简称“恒温扩增法”)联合腺苷脱氨酶(adenosine deaminase,ADA)、外周血结核感染 T 细胞斑点试验(T-SPOT.TB)检测对结核性胸膜炎患者早期诊断的价值。方法: 采用回顾性研究方法,选取2020年1月至2021年12月西安市胸科医院收治的不明原因胸腔积液患者427例作为研究对象。所有研究对象均进行胸腔积液恒温扩增法、胸腔积液ADA、胸腔积液结核分枝杆菌培养和外周血T-SPOT.TB检测。以临床综合诊断作为参考标准,评价恒温扩增法联合ADA、外周血T-SPOT.TB对结核性胸膜炎的早期诊断价值。结果: 427例研究对象中,279例(65.3%)确诊为结核性胸膜炎(结核性胸膜炎组),148例确诊为非结核性胸膜炎(非结核性胸膜炎组)。ADA的最佳诊断界值为19.5U/L。单一检测方法中,恒温扩增法的特异度为100.00%,敏感度为31.54%,与结核分枝杆菌培养类似(分别为100.00%和30.11%)。胸腔积液ADA和外周血T-SPOT.TB检测的敏感度分别为74.91%和84.95%,特异度分别为87.16%和74.91%。ADA+T-SPOT.TB并联、串联检测可以分别得到较高的敏感度(94.27%)和特异度(99.32%)。恒温扩增法、ADA、T-SPOT.TB 3种早期检测指标并联的检测效能与3种方法加结核分枝杆菌培养法并联检测一致。联合检测方法中ADA+T-SPOT.TB检测串联性价比最高,成本与效果的比值(C/E)为669.23。结论: 胸腔积液结核分枝杆菌DNA恒温扩增法与胸腔积液ADA、外周血T-SPOT.TB等检测方法联合可以为结核性胸膜炎患者早期诊断和治疗提供有力证据。

关键词: 分枝杆菌,结核, 胸膜炎, 核酸扩增技术, 腺苷脱氨酶, 临床实验室技术

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

中图分类号:

R521

赵国连, 雷倩, 王佩, 任云霞, 关靖. DNA实时荧光恒温扩增法联合检测对结核性胸膜炎的早期诊断价值[J]. 中国防痨杂志, 2023, 45(7): 681-686. doi: 10.19982/j.issn.1000-6621.20230055

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

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参考文献 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