IGI-LuNER: single-well multiplexed RT-qPCR test for SARS-CoV-2

Stahl, Elizabeth C.; Tsuchida, Connor A.; Hamilton, Jennifer R.; Lin-Shiao, Enrique; McDevitt, Shana L.; Moehle, Erica A.; Witkowsky, Lea B.; Tsui, C. Kimberly; Pestal, Kathleen; Gildea, Holly K.; McElroy, Matthew; Keller, Amanda; Sylvain, Iman; Williams, Clara; Hirsh, Ariana; Ciling, Alison; Ehrenberg, Alexander J.; Urnov, Fyodor D.; Ringeisen, Bradley R.; Giannikopoulos, Petros; Doudna, Jennifer A.

Commonly used RT-qPCR-based SARS-CoV-2 diagnostics require 2–3 separate reactions or rely on detection of a single viral target, adding time and cost or risk of false-negative results. Currently, no test combines detection of widely used SARS-CoV-2 E- and N-gene targets and a sample control in a single, multiplexed reaction. We developed the IGI-LuNER 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 (NER). This combined, cost-effective test can be performed in 384-well plates with detection sensitivity suitable for clinical reporting, and will aid in future sample pooling efforts, thus improving throughput of SARS-CoV-2 detection.