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用于快速诊断结核性淋巴结炎的环介导等温扩增(LAMP)检测:多靶点60分钟检测方法

Loop-mediated isothermal amplification (LAMP) assay for speedy diagnosis of tubercular lymphadenitis: The multi-targeted 60-minute approach.

作者信息

Sharma Megha, Sharma Kusum, Sharma Aman, Gupta Nalini, Rajwanshi Arvind

机构信息

Department of Medical Microbiology, Post Graduate Institute of Medical Education and Research, Chandigarh, India.

Department of Medical Microbiology, Post Graduate Institute of Medical Education and Research, Chandigarh, India.

出版信息

Tuberculosis (Edinb). 2016 Sep;100:114-117. doi: 10.1016/j.tube.2016.07.015. Epub 2016 Jul 29.

Abstract

INTRODUCTION

Tuberculous lymphadenitis (TBLA), the most common presentation of tuberculosis, poses a significant diagnostic challenge in the developing countries. Timely, accurate and cost-effective diagnosis can decrease the high morbidity associated with TBLA especially in resource-poor high-endemic regions. The loop-mediated isothermal amplification assay (LAMP), using two targets, was evaluated for the diagnosis of TBLA.

MATERIAL AND METHODS

LAMP assay using 3 sets of primers (each for IS6110 and MPB64) was performed on 170 fine needle aspiration samples (85 confirmed, 35 suspected, 50 control cases of TBLA). Results were compared against IS6110 PCR, cytology, culture and smear.

RESULTS

The overall sensitivity and specificity of LAMP assay, using multi-targeted approach, was 90% and 100% respectively in diagnosing TBLA. The sensitivity of multi-targeted LAMP, only MPB64 LAMP, only IS6110 LAMP and IS6110 PCR was 91.7%, 89.4%, 84.7% and 75.2%, respectively among confirmed cases and 85.7%, 77.1%, 68.5% and 60%, respectively among suspected cases of TBLA. Additional 12/120 (10%) cases were detected using multi-targeted method.

DISCUSSION

The multi-targeted LAMP, with its speedy and reliable results, is a potential diagnostic test for TBLA in low-resource countries.

摘要

引言

结核性淋巴结炎(TBLA)是结核病最常见的表现形式,在发展中国家构成了重大的诊断挑战。及时、准确且具有成本效益的诊断可以降低与TBLA相关的高发病率,尤其是在资源匮乏的高流行地区。使用两个靶标的环介导等温扩增检测法(LAMP)被用于TBLA的诊断评估。

材料与方法

使用3组引物(每组分别针对IS6110和MPB64)对170份细针穿刺样本(85例确诊、35例疑似、50例TBLA对照病例)进行LAMP检测。将结果与IS6110聚合酶链反应(PCR)检测、细胞学检查、培养及涂片检查结果进行比较。

结果

采用多靶标方法的LAMP检测在诊断TBLA时的总体敏感性和特异性分别为90%和100%。在确诊病例中,多靶标LAMP、仅MPB64的LAMP、仅IS6110的LAMP以及IS6110 PCR的敏感性分别为91.7%、89.4%、84.7%和75.2%;在疑似TBLA病例中,相应的敏感性分别为85.7%、77.1%、68.5%和60%。采用多靶标方法又检测出了额外的12/120(10%)例病例。

讨论

多靶标LAMP检测结果快速且可靠,是资源匮乏国家TBLA的一种潜在诊断方法。

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