Saliva TwoStep: An RT-LAMP saliva test for SARS-CoV-2 and its assessment in a large population
Saliva TwoStep: An RT-LAMP saliva test for SARS-CoV-2 and its assessment in a large population
Abstract 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) prepare saliva with heat and a stabilization solution, and 2) detect virus by incubating with a primer and enzyme mix at 65°C for 30 minutes. After incubation, saliva samples containing the SARS-CoV-2 genome turn bright yellow, while the negative samples remain pink. Because this test is pH dependent, it can react falsely to naturally acidic human saliva. Here, we report unique buffering conditions and protocols that rendered 295 healthy human saliva samples compatible with the test, producing zero false positives. We also evaluated the test on 168 saliva samples from individuals infected with SARS-CoV-2, and from 47 matched pairs of saliva and anterior nasal samples from infected individuals. The Saliva TwoStep test described here reliably identified infected individuals with 94% sensitivity and >99% specificity.
Barbachano-Guerrero Arturo、Decker Carolyn J.、Gonzales Patrick K.、Tat Kimngan L.、Ozeroff Christopher D.、McQueen Matthew B.、Leinwand Leslie、Parker Roy、Sawyer Sara L.、Clark Stephen K.、Wu Sharon S.、Muhlrad Denise、Meyerson Nicholas R.、Fattor Will T.、Brisson Gloria R.、Worden-Sapper Emma R.、Yang Qing、Saldi Tassa K.、Lasda Erika、Gilchrist Alison R.、Paige Camille L.、Hager Cole R.、Davis Jack C.、Fink Morgan R.
BioFrontiers Institute, University of Colorado Boulder, BoulderDepartment of Integrative Physiology, University of Colorado Boulder||Department of Biochemistry, University of Colorado BoulderBioFrontiers Institute, University of Colorado Boulder, BoulderBioFrontiers Institute, University of Colorado Boulder, BoulderBioFrontiers Institute, University of Colorado Boulder, BoulderDepartment of Integrative Physiology, University of Colorado BoulderBioFrontiers Institute, University of Colorado Boulder, Boulder||Department of Molecular, Cellular, and Developmental Biology, University of Colorado BoulderBioFrontiers Institute, University of Colorado Boulder, Boulder||Department of Molecular, Cellular, and Developmental Biology, University of Colorado Boulder||Department of Biochemistry, University of Colorado Boulder||Howard Hughes Medical Institute, University of Colorado BoulderBioFrontiers Institute, University of Colorado Boulder, Boulder||Department of Molecular, Cellular, and Developmental Biology, University of Colorado BoulderBioFrontiers Institute, University of Colorado Boulder, BoulderBioFrontiers Institute, University of Colorado Boulder, Boulder||Department of Molecular, Cellular, and Developmental Biology, University of Colorado BoulderDepartment of Integrative Physiology, University of Colorado Boulder||Department of Biochemistry, University of Colorado BoulderBioFrontiers Institute, University of Colorado Boulder, BoulderBioFrontiers Institute, University of Colorado Boulder, BoulderWardenburg Health Center, University of Colorado BoulderBioFrontiers Institute, University of Colorado Boulder, Boulder||Department of Molecular, Cellular, and Developmental Biology, University of Colorado BoulderBioFrontiers Institute, University of Colorado Boulder, Boulder||Department of Molecular, Cellular, and Developmental Biology, University of Colorado BoulderBioFrontiers Institute, University of Colorado Boulder, BoulderBioFrontiers Institute, University of Colorado Boulder, BoulderBioFrontiers Institute, University of Colorado Boulder, Boulder||Department of Molecular, Cellular, and Developmental Biology, University of Colorado BoulderBioFrontiers Institute, University of Colorado Boulder, BoulderBioFrontiers Institute, University of Colorado Boulder, BoulderDepartment of Molecular, Cellular, and Developmental Biology, University of Colorado BoulderBioFrontiers Institute, University of Colorado Boulder, Boulder
医学研究方法基础医学分子生物学
Barbachano-Guerrero Arturo,Decker Carolyn J.,Gonzales Patrick K.,Tat Kimngan L.,Ozeroff Christopher D.,McQueen Matthew B.,Leinwand Leslie,Parker Roy,Sawyer Sara L.,Clark Stephen K.,Wu Sharon S.,Muhlrad Denise,Meyerson Nicholas R.,Fattor Will T.,Brisson Gloria R.,Worden-Sapper Emma R.,Yang Qing,Saldi Tassa K.,Lasda Erika,Gilchrist Alison R.,Paige Camille L.,Hager Cole R.,Davis Jack C.,Fink Morgan R..Saliva TwoStep: An RT-LAMP saliva test for SARS-CoV-2 and its assessment in a large population[EB/OL].(2025-03-28)[2025-05-13].https://www.medrxiv.org/content/10.1101/2020.07.16.20150250.点此复制
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