Detection of SARS-CoV-2 RNA by multiplex RT-qPCR

  1. Eriko Kudo
  2. Benjamin Israelow
  3. Chantal B.F. Vogels
  4. Peiwen Lu
  5. Anne L. Wyllie
  6. Maria Tokuyama
  7. Arvind Venkataraman
  8. Doug E Brackney
  9. Isabel M. Ott
  10. Mary E. Petrone
  11. Rebecca Earnest
  12. Sarah Lapidus
  13. M. Catherine Muenker
  14. Adam J. Moore
  15. Arnau Casanovas-Massana
  16. Yale IMPACT Research Team
  17. Saad B. Omer
  18. Charles S. Dela Cruz
  19. Shelli F. Farhadian
  20. Albert I. Ko
  21. Nathan D. Grubaugh
  22. Akiko Iwasaki
2 evaluations Published on
Read the full article Related papers
This article on Sciety

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.

Related articles

Related articles are currently not available for this article.