Development of a Saliva-Optimized RT-LAMP Assay for SARS-CoV-2

Albert D Yu; Kristina Galatsis; Jian Zheng; Jasmine Quynh Le; Dingbang Ma; Stanley Perlman; Michael Rosbash

doi:10.7171/jbt.21-3203-005

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Development of a Saliva-Optimized RT-LAMP Assay for SARS-CoV-2

Journal article

Open access

Peer reviewed

Development of a Saliva-Optimized RT-LAMP Assay for SARS-CoV-2

Albert D Yu, Kristina Galatsis, Jian Zheng, Jasmine Quynh Le, Dingbang Ma, Stanley Perlman and Michael Rosbash

Journal of biomolecular techniques, Vol.32(3), pp.102-113

09/2021

DOI: 10.7171/jbt.21-3203-005

PMCID: PMC8730519

PMID: 35027868

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Abstract

Conventional reverse transcription quantitative polymerase chain reaction (RT-qPCR) technology has struggled to fulfill the unprecedented need for diagnostic testing created by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic. Complexity and cost hinder access to testing, and long turnaround time decreases its utility. To ameliorate these issues, we focus on saliva and introduce several advances to colorimetric reverse transcription loop-mediated isothermal amplification (RT-LAMP) technology; RT-LAMP offers a minimal equipment alternative to RT-qPCR. First, we validated the use of the novel dye LAMPShade Violet (LSV), which improves the visual clarity and contrast of the colorimetric readout. Second, we compared different inactivation conditions on infectivity and RNA yield from saliva. Third, we developed a 10-minute RNA purification protocol from saliva. We call this magnetic bead protocol . Finally, we developed a magnetic stick, StickLAMP, which provides reliable bead-based RNA purification as well as simple and low-cost access to scalable testing from saliva.

COVID-19

saliva

SARS-CoV-2

magnetic beads

coronavirus

loop-mediated isothermal amplification

Details

Title: Subtitle: Development of a Saliva-Optimized RT-LAMP Assay for SARS-CoV-2
Creators: Albert D Yu - Brandeis University
Kristina Galatsis - Brandeis University
Jian Zheng - University of Iowa, Microbiology and Immunology
Jasmine Quynh Le - Brandeis University
Dingbang Ma - Brandeis University
Stanley Perlman - University of Iowa, Microbiology and Immunology
Michael Rosbash - Howard Hughes Medical Institute and Department of Biology, Brandeis University, Waltham, Massachusetts, USA
Resource Type: Journal article
Publication Details: Journal of biomolecular techniques, Vol.32(3), pp.102-113
DOI: 10.7171/jbt.21-3203-005
PMID: 35027868
PMCID: PMC8730519
eISSN: 1943-4731
Language: English
Date published: 09/2021
Academic Unit: Microbiology and Immunology; Stead Family Department of Pediatrics; Iowa Neuroscience Institute; Infectious Disease (Pediatrics)
Record Identifier: 9984209047702771

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