Open AccessLuNER: Multiplexed SARS-CoV-2 detection in clinical swab and wastewater samples
Author(s) -
Elizabeth C. Stahl,
Allan Gopez,
Connor A. Tsuchida,
Vinson B. Fan,
Erica A. Moehle,
Lea B. Witkowsky,
Jennifer Hamilton,
Enrique Lin-Shiao,
Matthew McElroy,
Shana L. McDevitt,
Alison Ciling,
C. Kimberly Tsui,
Kathleen Pestal,
Holly K. Gildea,
Amanda Keller,
Iman Sylvain,
Clara Williams,
Ariana Hirsh,
Alexander J. Ehrenberg,
Rose Kantor,
Matthew Metzger,
Kara L. Nelson,
Fyodor D. Urnov,
Bradley R. Ringeisen,
Petros Giannikopoulos,
Jennifer A. Doudna
Publication year - 2021
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0258263
Subject(s) - pooling , covid-19 , molecular diagnostics , computational biology , virology , computer science , biology , medicine , bioinformatics , artificial intelligence , disease , pathology , outbreak , infectious disease (medical specialty)
Clinical and surveillance testing for the SARS-CoV-2 virus relies overwhelmingly on RT-qPCR-based diagnostics, yet several popular assays require 2–3 separate reactions or rely on detection of a single viral target, which adds significant time, cost, and risk of false-negative results. Furthermore, multiplexed RT-qPCR tests that detect at least two SARS-CoV-2 genes in a single reaction are typically not affordable for large scale clinical surveillance or adaptable to multiple PCR machines and plate layouts. We developed a RT-qPCR assay using the Luna Probe Universal One-Step RT-qPCR master mix with publicly available primers and probes to detect SARS-CoV-2 N gene, E gene, and human RNase P (LuNER) to address these shortcomings and meet the testing demands of a university campus and the local community. This cost-effective test is compatible with BioRad or Applied Biosystems qPCR machines, in 96 and 384-well formats, with or without sample pooling, and has a detection sensitivity suitable for both clinical reporting and wastewater surveillance efforts.