Abstract
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.
Highlights
Diagnostic testing remains a vital component of the public health response to the global pandemic caused by the SARS-CoV-2 [1,2,3,4,5,6,7]
Quantitative reverse transcription polymerase chain reaction (RT-qPCR) of RNA extracted from nasal swabs is the most common diagnostic testing method to detect SARS-CoV-2 [8, 9]
The UC Berkeley Committee for Protection of Human Subjects determined that all the analyses presented in this manuscript do not qualify as human subjects research as the data sets were de-identified to those analyzing them for these results (IRB submission # 2020-04-13177)
Summary
Diagnostic testing remains a vital component of the public health response to the global pandemic caused by the SARS-CoV-2 [1,2,3,4,5,6,7]. Quantitative reverse transcription polymerase chain reaction (RT-qPCR) of RNA extracted from nasal swabs is the most common diagnostic testing method to detect SARS-CoV-2 [8, 9]. Current tests rely on the detection of genomic or sub-genomic viral RNA corresponding to the envelope (E) gene, the nucleocapsid (N) gene, the spike (S) gene, and/or the ORF1ab locus [10,11,12], as well as suitable controls. Variability in test format as well as reagent availability, quality, and cost continue to hamper widespread use for both symptomatic and asymptomatic testing.
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
