eLife (Mar 2021)
Saliva TwoStep for rapid detection of asymptomatic SARS-CoV-2 carriers
- Qing Yang,
- Nicholas R Meyerson,
- Stephen K Clark,
- Camille L Paige,
- Will T Fattor,
- Alison R Gilchrist,
- Arturo Barbachano-Guerrero,
- Benjamin G Healy,
- Emma R Worden-Sapper,
- Sharon S Wu,
- Denise Muhlrad,
- Carolyn J Decker,
- Tassa K Saldi,
- Erika Lasda,
- Patrick Gonzales,
- Morgan R Fink,
- Kimngan L Tat,
- Cole R Hager,
- Jack C Davis,
- Christopher D Ozeroff,
- Gloria R Brisson,
- Matthew B McQueen,
- Leslie A Leinwand,
- Roy Parker,
- Sara L Sawyer
Affiliations
- Qing Yang
- ORCiD
- BioFrontiers Institute, University of Colorado Boulder, Boulder, United States; Department of Molecular, Cellular and Developmental Biology, University of Colorado Boulder, Boulder, United States
- Nicholas R Meyerson
- BioFrontiers Institute, University of Colorado Boulder, Boulder, United States; Darwin Biosciences Inc, Boulder, United States
- Stephen K Clark
- BioFrontiers Institute, University of Colorado Boulder, Boulder, United States; Darwin Biosciences Inc, Boulder, United States
- Camille L Paige
- ORCiD
- BioFrontiers Institute, University of Colorado Boulder, Boulder, United States
- Will T Fattor
- ORCiD
- BioFrontiers Institute, University of Colorado Boulder, Boulder, United States
- Alison R Gilchrist
- BioFrontiers Institute, University of Colorado Boulder, Boulder, United States; Department of Molecular, Cellular and Developmental Biology, University of Colorado Boulder, Boulder, United States
- Arturo Barbachano-Guerrero
- ORCiD
- BioFrontiers Institute, University of Colorado Boulder, Boulder, United States
- Benjamin G Healy
- Department of Mechanical Engineering, University of Colorado Boulder, Boulder, United States
- Emma R Worden-Sapper
- ORCiD
- BioFrontiers Institute, University of Colorado Boulder, Boulder, United States; Department of Molecular, Cellular and Developmental Biology, University of Colorado Boulder, Boulder, United States
- Sharon S Wu
- BioFrontiers Institute, University of Colorado Boulder, Boulder, United States; Department of Molecular, Cellular and Developmental Biology, University of Colorado Boulder, Boulder, United States; Interdisciplinary Quantitative Biology Graduate Program, University of Colorado Boulder, Boulder, United States
- Denise Muhlrad
- Department of Biochemistry, University of Colorado Boulder, Boulder, United States; Howard Hughes Medical Institute, University of Colorado Boulder, Boulder, United States
- Carolyn J Decker
- Department of Biochemistry, University of Colorado Boulder, Boulder, United States; Howard Hughes Medical Institute, University of Colorado Boulder, Boulder, United States
- Tassa K Saldi
- BioFrontiers Institute, University of Colorado Boulder, Boulder, United States
- Erika Lasda
- BioFrontiers Institute, University of Colorado Boulder, Boulder, United States
- Patrick Gonzales
- BioFrontiers Institute, University of Colorado Boulder, Boulder, United States; Department of Integrative Physiology, University of Colorado Boulder, Boulder, United States
- Morgan R Fink
- ORCiD
- BioFrontiers Institute, University of Colorado Boulder, Boulder, United States
- Kimngan L Tat
- BioFrontiers Institute, University of Colorado Boulder, Boulder, United States
- Cole R Hager
- BioFrontiers Institute, University of Colorado Boulder, Boulder, United States
- Jack C Davis
- Department of Molecular, Cellular and Developmental Biology, University of Colorado Boulder, Boulder, United States
- Christopher D Ozeroff
- BioFrontiers Institute, University of Colorado Boulder, Boulder, United States
- Gloria R Brisson
- Wardenburg Health Center, University of Colorado Boulder, Boulder, United States
- Matthew B McQueen
- Department of Integrative Physiology, University of Colorado Boulder, Boulder, United States
- Leslie A Leinwand
- ORCiD
- BioFrontiers Institute, University of Colorado Boulder, Boulder, United States; Department of Molecular, Cellular and Developmental Biology, University of Colorado Boulder, Boulder, United States
- Roy Parker
- ORCiD
- BioFrontiers Institute, University of Colorado Boulder, Boulder, United States; Department of Molecular, Cellular and Developmental Biology, University of Colorado Boulder, Boulder, United States; Department of Biochemistry, University of Colorado Boulder, Boulder, United States; Howard Hughes Medical Institute, University of Colorado Boulder, Boulder, United States
- Sara L Sawyer
- ORCiD
- BioFrontiers Institute, University of Colorado Boulder, Boulder, United States; Department of Molecular, Cellular and Developmental Biology, University of Colorado Boulder, Boulder, United States
- DOI
- https://doi.org/10.7554/eLife.65113
- Journal volume & issue
-
Vol. 10
Abstract
Here, we develop a simple molecular test for SARS-CoV-2 in saliva based on reverse transcription loop-mediated isothermal amplification. 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.
Keywords
