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利用环介导等温扩增(LAMP)试验直接从呼吸道样本中快速检测 SARS-CoV-2 RNA。

Fast Detection of SARS-CoV-2 RNA Directly from Respiratory Samples Using a Loop-Mediated Isothermal Amplification (LAMP) Test.

机构信息

Institute for Virology, University Hospital Essen, University of Duisburg-Essen, 45147 Essen, Germany.

Labor Dr. Krause und Kollegen MVZ GmbH, 24106 Kiel, Germany.

出版信息

Viruses. 2021 Apr 29;13(5):801. doi: 10.3390/v13050801.

Abstract

The availability of simple SARS-CoV-2 detection methods is crucial to contain the COVID-19 pandemic. This study examined whether a commercial LAMP assay can reliably detect SARS-CoV-2 genomes directly in respiratory samples without having to extract nucleic acids (NA) beforehand. Nasopharyngeal swabs (NPS, = 220) were tested by real-time reverse transcription (RT)-PCR and with the LAMP assay. For RT-PCR, NA were investigated. For LAMP, NA from 26 NPS in viral transport medium (VTM) were tested. The other 194 NPS were analyzed directly without prior NA extraction (140 samples in VTM; 54 dry swab samples stirred in phosphate buffered saline). Ten NPS were tested directly by LAMP using a sous-vide cooking unit. The isothermal assay demonstrated excellent specificity (100%) but moderate sensitivity (68.8%), with a positive predictive value of 1 and a negative predictive value of 0.65 for direct testing of NPS in VTM. The use of dry swabs, even without NA extraction, improved the analytical sensitivity; up to 6% of samples showed signs of inhibition. LAMP could be performed successfully with a sous-vide cooking unit. This technique is very fast, requires little laboratory resources, and can replace rapid antigen tests or verify reactive rapid tests on-site.

摘要

简便的 SARS-CoV-2 检测方法对于控制 COVID-19 大流行至关重要。本研究旨在探讨一种商用 LAMP 检测方法是否可以直接从呼吸道样本中可靠地检测到 SARS-CoV-2 基因组,而无需事先提取核酸(NA)。采用实时 RT-PCR 和 LAMP 检测法对鼻咽拭子(NPS,n=220)进行了检测。对于 RT-PCR,检测了 NA。对于 LAMP,检测了 26 份置于病毒运输介质(VTM)中的 NPS 的 NA。其余 194 份 NPS 未经 NA 提取直接进行分析(140 份 VTM 样本;54 份干拭子样本在磷酸盐缓冲盐水(PBS)中搅拌)。10 份 NPS 直接使用水浴烹饪设备进行 LAMP 检测。等温检测法显示出极好的特异性(100%),但敏感性中等(68.8%),对 VTM 中直接检测 NPS 的阳性预测值为 1,阴性预测值为 0.65。即使不提取 NA,使用干拭子也可提高分析灵敏度;多达 6%的样本显示出抑制迹象。LAMP 可以成功地使用水浴烹饪设备进行。该技术非常快速,所需实验室资源较少,可替代快速抗原检测,或在现场验证反应性快速检测。

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