Abstract
The current RT-qPCR assay recommended for SARS-CoV-2 testing in the United States requires analysis of three genomic targets per sample: two viral and one host. To simplify testing and reduce the volume of required reagents, we developed 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 CDC, 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 singleplex assay adapted for research purposes. Low copies (>500 copies / reaction) of SARS-CoV-2 RNA were consistently detected by the multiplex RT-qPCR. Our novel multiplex RT-qPCR improves upon current singleplex diagnostics by saving reagents, costs, time and labor.
Competing Interest Statement
The authors have declared no competing interest.
Footnotes
↵A Research was supported in part by the National Institute of Health grants, 2T32AI007517-16 (to BI), AI054359 and AI127429 (to AI), the Japan Society for the Promotion of Science (to EK), Women’s Health Research at Yale Pilot Project Program, Emergent Ventures at the Mercatus Center, George Mason University, Mathers Foundation and the Ludwig Family Foundation. A.I. is an Investigator of the Howard Hughes Medical Institute.
↵B None of the authors have any conflicts of interest to declare.
