Diagnostic value of loop-mediated isothermal amplification method in non sputum or smear negative pulmonary tuberculosis

doi:10.19983/j.issn.2096-8493.20230001

Abstract

Abstract:

Objective: To explore the diagnostic value of loop-mediated isothermal amplification (LAMP) in detecting bronchoalveolar lavage fluid (BALF) in non sputum or smear negative pulmonary tuberculosis. Methods: The prospective study method was adopted, with the subjects being the sputum free or smear negative suspected pulmonary tuberculosis patients who were admitted to Wushan County People’s Hospital of Chongqing from April 2021 to April 2022 and met the inclusion criteria; they all brushed under bronchoscope for fluorescent staining microscopy (referred to as “smear”), and took BALF for LAMP and GeneXpert MTB/RIF (referred to as “Xpert”) to detect Mycobacterium tuberculosis complex DNA. With the final clinical diagnosis as the reference standard, the detection efficiency of the three methods was analyzed. Results: A total of 232 patients were included in the study. The patients were divided into smear negative/sputum free pulmonary tuberculosis patients (139 cases) and non tuberculosis pulmonary disease patients (93 cases) according to the final clinical diagnosis. The positive rates of LAMP, Xpert and smear in 232 patients with suspected pulmonary tuberculosis were 43.53% (101/232), 43.97% (102/232) and 8.19% (19/232), respectively. Taking the final clinical diagnosis as the reference standard, the detection sensitivity of LAMP, Xpert and smear for sputum free or smear negative pulmonary tuberculosis were 71.94% (100/139), 72.66% (101/139) and 13.67% (19/139) respectively; the specificity were 98.92% (92/93), 98.92% (92/93) and 100.00% (93/93) respectively; the diagnostic coincidence rates were 82.76% (192/232), 83.19% (193/232) and 48.28% (112/232); the Yoden index were 0.709, 0.716 and 0.137, respectively. Conclusion: The detection efficiency of LAMP for BALF in patients with sputum free or smear negative pulmonary tuberculosis is basically consistent with that of Xpert, significantly improving the early diagnosis rate; in addition, LAMP is easy to operate and cheap, which is more suitable for the promotion and application of grass-roots medical institutions.

Key words: Tuberculosis, pulmonary, Bronchoalveolar lavage fluid, Nucleic acid amplification techniques, Evaluation studies

CLC Number:

R521

Zeng Fang, Zhu Xiaoxia, Gong Rui, Liu Chuanhong, Shi Dejun, Zhao Ling, Yang Song. Diagnostic value of loop-mediated isothermal amplification method in non sputum or smear negative pulmonary tuberculosis[J]. Journal of Tuberculosis and Lung Disease , 2023, 4(1): 66-71. doi: 10.19983/j.issn.2096-8493.20230001

Figures/Tables 2

References 24

[1]	World Health Organization.Global tuberculosis report 2021. Geneva: World Health Organization, 2021.
[2]	World Health Organization.Global tuberculosis report 2022. Geneva: World Health Organization, 2022.
[3]	中华人民共和国国务院办公厅. 国务院办公厅关于印发“十三五”全国结核病防治规划的通知. 国办发〔2017〕16号. 2017-02-01.
[4]	中华人民共和国国家卫生和计划生育委员会. WS 288—2017肺结核诊断2017-11-09.
[5]	国家感染性疾病临床医学研究中心, 深圳市第三人民医院,《中国防痨杂志》编辑委员会. 肺结核活动性判断规范及临床应用专家共识. 中国防痨杂志, 2020, 42(4):301-307. doi:10.3969/j.issn.1000-6621.2020.04.001. doi: 10.3969/j.issn.1000-6621.2020.04.001.
[6]	中华人民共和国国家卫生和计划生育委员会. WS 196—2017结核病分类. 2017-11-09.
[7]	中华医学会结核病学分会临床检验专业委员会. 结核病病原学分子诊断专家共识. 中华结核和呼吸杂志, 2018, 41(9):688-695. doi:10.3760/cma.j.issn.100l-0939.2018.09.008. doi: 10.3760/cma.j.issn.100l-0939.2018.09.008.
[8]	王雅娟, 曹新益, 刘升明. 宏基因组二代测序和GeneXpert MTB/RIF对结核病诊断价值的Meta分析. 中国防痨杂志, 2022, 44(12):1327-1337. doi:10.19982/j.issn.1000-6621.20220303. doi: 10.19982/j.issn.1000-6621.20220303.
[9]	彭荣, 龚倩. GeneXpert MTB/RIF检测肺泡灌洗液对无痰或少痰肺结核的诊断价值. 检验医学与临床, 2020, 17(22):3267-3269. doi:10.3969/j.issn.1672-9455.2020.22.012. doi: 10.3969/j.issn.1672-9455.2020.22.012.
[10]	Li Y, Fan P, Zhou S, et al. Loop-mediated isothermal amplification (LAMP): a novel rapid detection platform for pathogens. Microb Pathog, 2017, 107: 54-61. doi:10.1016/j.micpath.2017.03.016. doi: 10.1016/j.micpath.2017.03.016. URL
[11]	戴广明, 曹以诚, 杜正平, 等. 结核分枝杆菌环介导恒温扩增(LAMP)快速检测方法的评价. 中国防痨杂志, 2011, 33(1):47-51.
[12]	陈钰, 罗力涵, 李莎, 等. 结核分枝杆菌 LAMP 检测方法的建立及应用. 中国国境卫生检疫杂志, 2017, 40(4):242-247. doi:10.16408/j.1004-9770.2017.04.004. doi: 10.16408/j.1004-9770.2017.04.004.
[13]	李秀萍, 赵雪梅, 严晓芸, 等. T-SPOT.TB、TB-LAMP、TB-DNA定量对肺结核的诊断价值分析. 湖南师范大学学报(医学版), 2021, 18(2):185-188. doi:10.3969/j.issn.1673-016X.2021.02.053. doi: 10.3969/j.issn.1673-016X.2021.02.053.
[14]	余欢成, 陈玲, 祝国强, 等. 支气管肺泡灌洗液结核杆菌检测在菌阴肺结核中的诊断价值. 中国现代医生, 2018, 56(12):73-75,79.
[15]	陈霞, 张建勇, 赵建军. 支气管肺泡灌洗液应用TB-LAMP对菌阴肺结核的诊断价值. 中国感染控制杂志, 2020, 19(7):643-647. doi:12138/j.issn.1671-9638.20205739. doi: 12138/j.issn.1671-9638.20205739.
[16]	Yu G, Shen Y, Zhong F, et al. Diagnostic accuracy of the loop-mediated isothermal amplification assay for extrapulmonary tuberculosis: a meta-analysis. PLoS One, 2018, 13(6): e0199290. doi:10.1371/journal.pone.0199290. doi: 10.1371/journal.pone.0199290. URL
[17]	廖洁, 陈小林, 李小玲. LAMP在临床上诊断肺结核的应用评价. 当代医学, 2021, 27(7): 143-144. doi:10.3969/j.issn.1009-4393.2021.07.060. doi: 10.3969/j.issn.1009-4393.2021.07.060.
[18]	刘亚芹, 陈焕民, 陈鸣. LAMP技术检测支气管肺泡灌洗液在疑似肺结核中的诊断价值. 标记免疫分析与临床, 2020, 27(6):1037-1039. doi:10.11748/bjmy.issn.1006-1703.2020.06.026. doi: 10.11748/bjmy.issn.1006-1703.2020.06.026.
[19]	Fan L, Guan B, Cheng M, et al. A Comprehensive Evaluation of a Loop-Mediated Isothermal Amplification Assay for the Diagnosis of Pulmonary Tuberculosis in Children Using Bronchoalveolar Lavage Fluid. Infect Drug Resist, 2022, 15:975-987. doi:10.2147/IDR.S354660. doi: 10.2147/IDR.S354660 pmid: 35299853
[20]	欧维正, 刘海兰, 雷毅娜, 等. 支气管肺泡灌洗液涂片抗酸染色、L-J培养、TB-LAMP、Xpert MTB/RIF检测在肺结核诊断中的应用比较. 山东医药, 2020, 60(23):57-60. doi:10.3969/j.issn.1002-266X.2020.23.015. doi: 10.3969/j.issn.1002-266X.2020.23.015.
[21]	贾枫, 叶海容, 曾云龙, 等. 环介导等温扩增技术(LAMP)在肺结核诊断中的应用研究. 实验与检验医学, 2021, 39(5):1041-1043. doi:10.3969/j.issn.1674-1129.2021.05.001. doi: 10.3969/j.issn.1674-1129.2021.05.001.
[22]	Wang Z, Zang Y, Gao Y, et al. Evaluation of bronchoalveolar lavage fluid combined with the loop-mediated isothermal amplification assay in lower respiratory tract infections. Am J Transl Res, 2020, 12(7):4009-4016. pmid: 32774754
[23]	Kim CK, Cho EA, Shin DM, et al. Comparative evaluation of the loop-mediat ed isothermal amplification assay for detecting pulmonary tuberculosis. Ann Lab Med, 2018, 38(2): 119-124. doi:10.3343/alm.2018.38.2.119. doi: 10.3343/alm.2018.38.2.119. URL
[24]	Shete PB, Farr K, Strnad L, et al. Diagnostic accuracy of TB-LAMP for pulmonary tuberculosis: a systematic review and meta-analysis. BMC Infect Dis, 2019, 19(1):268. doi:10.1186/s12879-019-3881-y. doi: 10.1186/s12879-019-3881-y pmid: 30890135

LAMP检测	Xpert检测		合计(例)	χ²值	P值
LAMP检测	阳性(例)	阴性(例)	合计(例)	χ²值	P值
阳性	94	6	100	0.077	0.782
阴性	7	32	39
合计	101	38	139

检测方法	临床诊断(例)		敏感度 (%)	特异度 (%)	阳性预测值 (%)	阴性预测值 (%)	一致率 (%)	约登指数
检测方法	肺结核	非活动性结核性肺病	敏感度 (%)	特异度 (%)	阳性预测值 (%)	阴性预测值 (%)	一致率 (%)	约登指数
LAMP			71.94	98.92	99.01	70.23	82.76	0.709
阳性	100	1
阴性	39	92
Xpert			72.66	98.92	99.02	70.23	83.19	0.716
阳性	101	1
阴性	39	92
涂片镜检			13.67	100.00	100.00	43.66	48.28	0.137
阳性	19	0
阴性	120	93