Year: 2020
Type: article
Abstract: The current quantitative reverse transcription PCR (RT-qPCR) assay recommended for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) testing in the United States requires analysis of 3 genomic targets per sample: 2 viral and 1 host. To simplify testing and reduce the volume of required reagents, we devised a multiplex RT-qPCR assay to detect SARS-CoV-2 in a single reaction. We used existing N1, N2, and RP primer and probe sets by the Centers for Disease Control and Prevention, but substituted fluorophores to allow multiplexing of the assay. The cycle threshold (Ct) values of our multiplex RT-qPCR were comparable to those obtained by the single assay adapted for research purposes. Low copy numbers (≥500 copies/reaction) of SARS-CoV-2 RNA were consistently detected by the multiplex RT-qPCR. Our novel multiplex RT-qPCR improves upon current single diagnostics by saving reagents, costs, time, and labor.
Source: PLoS Biology
Authors: Eriko Kudo, Benjamin Israelow, Chantal B.F. Vogels, Peiwen Lu, Anne L. Wyllie, Maria Tokuyama, Arvind Venkataraman, Doug E Brackney, Isabel M. Ott, Mary E. Petrone, Rebecca Earnest, Sarah Lapidus, M. Catherine Muenker, Adam J. Moore, Arnau Casanovas-Massana, Saad B. Omer, Charles S. Dela Cruz, Shelli F. Farhadian, Albert I. Ko, Nathan D. Grubaugh, Akiko Iwasaki
Language: English
Subfield: Infectious Diseases
Field: Medicine
Domain: Health Sciences
Open Access status: gold
Estimated APC paid: $3,000
Funders: Howard Hughes Medical Institute, G. Harold and Leila Y. Mathers Charitable Foundation, Yale University, George Mason University, Strong, Ludwig Family Foundation, National Institutes of Health, Mercatus Center, George Mason University, Japan Society for the Promotion of Science, National Institute of Allergy and Infectious Diseases, National Center for Advancing Translational Sciences
OpenAlex ID: W3092279081