Yang Qing, Meyerson Nicholas R, Clark Stephen K, Paige Camille L, Fattor Will T, Gilchrist Alison R, Barbachano-Guerrero Arturo, Healy Benjamin G, Worden-Sapper Emma R, Wu Sharon S, Muhlrad Denise, Decker Carolyn J, Saldi Tassa K, Lasda Erika, Gonzales Patrick K, Fink Morgan R, Tat Kimngan L, Hager Cole R, Davis Jack C, Ozeroff Christopher D, Brisson Gloria R, McQueen Matthew B, Leinwand Leslie, Parker Roy, Sawyer Sara L
BioFrontiers Institute, University of Colorado Boulder, Boulder, Colorado, 80303.
Department of Molecular, Cellular, and Developmental Biology, University of Colorado Boulder, Boulder, Colorado, 80303.
medRxiv. 2021 Feb 16:2020.07.16.20150250. doi: 10.1101/2020.07.16.20150250.
Here, we develop a simple molecular test for SARS-CoV-2 in saliva based on reverse transcription loop-mediated isothermal amplification (RT-LAMP). The test has two steps: 1) heat saliva with a stabilization solution, and 2) detect virus by incubating with a primer/enzyme mix. After incubation, saliva samples containing the SARS-CoV-2 genome turn bright yellow. Because this test is pH dependent, it can react falsely to some naturally acidic saliva samples. We report unique saliva stabilization protocols that rendered 295 healthy saliva samples compatible with the test, producing zero false positives. We also evaluated the test on 278 saliva samples from individuals who were infected with SARS-CoV-2 but had no symptoms at the time of saliva collection, and from 54 matched pairs of saliva and anterior nasal samples from infected individuals. The Saliva TwoStep test described herein identified infections with 94% sensitivity and >99% specificity in individuals with sub-clinical (asymptomatic or pre-symptomatic) infections.
在此，我们基于逆转录环介导等温扩增技术（RT-LAMP）开发了一种用于检测唾液中新型冠状病毒（SARS-CoV-2）的简单分子检测方法。该检测方法有两个步骤：1）将唾液与稳定溶液一起加热；2）通过与引物/酶混合物孵育来检测病毒。孵育后，含有SARS-CoV-2基因组的唾液样本会变成亮黄色。由于该检测方法依赖于pH值，它可能会对一些天然酸性的唾液样本产生假反应。我们报告了独特的唾液稳定方案，该方案使295份健康唾液样本与该检测方法兼容，未产生假阳性结果。我们还对278份来自感染了SARS-CoV-2但在采集唾液时无症状的个体的唾液样本，以及54对来自感染个体的匹配唾液和前鼻样本进行了检测评估。本文所述的唾液两步检测法在亚临床（无症状或症状前）感染个体中检测感染的灵敏度为94%，特异性>99%。
