使用PURE TB-LAMP检测法快速检测痰标本中的结核分枝杆菌复合群

Rapid detection of Mycobacterium tuberculosis complex in sputum Samples using PURE TB-LAMP assay.

作者信息

N'guessan K, Horo K, Coulibaly I, Adegbele J, Kouame-Adjei N, Seck-Angu H, Guei A, Kouakou J, Dosso M

机构信息

Laboratoire Nationale de Référence de la Tuberculose, Institut Pasteur de Côte d'Ivoire, Abidjan, Cote d'Ivoire.

Service de Pneumologie CHU de Cocody, Abidjan, Cote d'Ivoire.

出版信息

Int J Mycobacteriol. 2016 Dec;5 Suppl 1:S164-S165. doi: 10.1016/j.ijmyco.2016.09.025. Epub 2016 Nov 9.

DOI:10.1016/j.ijmyco.2016.09.025

PMID:28043530

Abstract

OBJECTIVE/BACKGROUND: Lack of rapid and accurate diagnostic testing is a critical obstacle to global tuberculosis (TB) control. Sensitivity of sputum smear microscopy (SSM) is not optimal; however, it remains the most prevalent tool for TB confirmation in poor countries. As a part of passive case finding of TB detection, this study was conducted to determine the clinical performance of PURE TB-LAMP assay using liquid culture medium as the gold standard.

METHODS

Centre Antituberculeux de Yopougon is one of the 17 intermediate Tuberculosis centers in Côte d'Ivoire. A standardized questionnaire was submitted to patients with signs and symptoms consistent with tuberculosis by a trained caregiver. After obtaining signed consent forms, sputum samples were collected according to National TB Control Programme guidelines (spot-morning). SSM after Ziehl-Neelsen staining and TB-LAMP assay were blindly performed on the first sample. Samples transported to Institut Pasteur de Côte d'Ivoire were decontaminated according to the N-acetyl-L-Cystein method. In Mycobacteria Growth Indicator Tube (MGIT), 500mL of pellets were inoculated and incubated in the MGIT 960 system. MPT64 antigen was detected in positive cultures.

RESULTS

Of the 500 patients enrolled, 469 (232men and 239 women) patients were included. The mean ages of men and women were 36.9 (15-86) and 37.3 (15-37.3) years, respectively. There were 56 (12.2%) HIV-infected patients, including 14 women. Clinical isolates of M. tuberculosis complex were detected for 157 (33.5%) patients. Compared with culturing, the overall sensitivity and specificity of SSM were 86% (95% confidence interval [CI]=81-91) and 96% (95% CI=94-98), respectively. The overall sensitivity and specificity for TB-LAMP was 92% (95% CI=0.88-0.96) and 94% (95% CI=0.91-0.97), respectively. Positive likelihood ratios for TB-LAMP and SSM were 15.3 and 21.5, respectively, and negative likelihood ratios for TB-LAMP and SSM were 0.09 and 0.15, respectively. Among the 469 patients, active tuberculosis was detected using TB-LAMP assay and SSM in 162 (34.5%) and 147 (31.3%) patients, respectively.

CONCLUSION

For accurate diagnostic of pulmonary TB, TB-LAMP could be used as a tool of the first intention.

摘要

目的/背景：缺乏快速准确的诊断检测是全球结核病控制的关键障碍。痰涂片显微镜检查（SSM）的敏感性并不理想；然而，在贫穷国家，它仍是结核病确诊最常用的工具。作为结核病检测被动病例发现的一部分，本研究以液体培养基作为金标准，旨在确定PURE TB-LAMP检测法的临床性能。

方法

约普贡抗结核中心是科特迪瓦的17个中级结核病中心之一。一名经过培训的护理人员向有结核病相关体征和症状的患者发放标准化问卷。在获得签署的同意书后，按照国家结核病控制规划指南（即时痰-晨痰）采集痰样本。对首个样本进行萋尼氏染色后的SSM和TB-LAMP检测，检测过程均采用盲法。运送到科特迪瓦巴斯德研究所的样本按照N-乙酰-L-半胱氨酸法进行净化处理。在分枝杆菌生长指示管（MGIT）中接种500mL沉淀，并在MGIT 960系统中进行培养。在阳性培养物中检测MPT64抗原。

结果

纳入的500例患者中，469例（232例男性和239例女性）被纳入研究。男性和女性的平均年龄分别为36.9岁（15 - 86岁）和37.3岁（15 - 37.3岁）。有56例（12.2%）HIV感染患者，其中包括14例女性。157例（33.5%）患者检测到结核分枝杆菌复合群的临床分离株。与培养法相比，SSM的总体敏感性和特异性分别为86%（95%置信区间[CI]=81 - 91）和96%（95% CI=94 - 98）。TB-LAMP的总体敏感性和特异性分别为92%（95% CI=0.88 - 0.96）和94%（95% CI=0.91 - 0.97）。TB-LAMP和SSM的阳性似然比分别为15.3和21.5，阴性似然比分别为0.09和0.15。在469例患者中，分别有162例（34.5%）和147例（31.3%）患者通过TB-LAMP检测法和SSM检测到活动性结核病。