PCR assay to enhance global surveillance for SARS-CoV-2 variants of concern.
Vogels CBF., Breban MI., Alpert T., Petrone ME., Watkins AE., Ott IM., de Jesus JG., Claro IM., Ferreira GM., Crispim MAE., Brazil-UK CADDE Genomic Network None., Singh L., Tegally H., Anyaneji UJ., NGS-SA None., Hodcroft EB., Mason CE., Khullar G., Metti J., Dudley JT., MacKay MJ., Nash M., Wang J., Liu C., Hui P., Murphy S., Neal C., Laszlo E., Landry ML., Muyombwe A., Downing R., Razeq J., de Oliveira T., Faria NR., Sabino EC., Neher RA., Fauver JR., Grubaugh ND.
With the emergence of SARS-CoV-2 variants that may increase transmissibility and/or cause escape from immune responses 1-3 , there is an urgent need for the targeted surveillance of circulating lineages. It was found that the B.1.1.7 (also 501Y.V1) variant first detected in the UK 4,5 could be serendipitously detected by the ThermoFisher TaqPath COVID-19 PCR assay because a key deletion in these viruses, spike Δ69-70, would cause a "spike gene target failure" (SGTF) result. However, a SGTF result is not definitive for B.1.1.7, and this assay cannot detect other variants of concern that lack spike Δ69-70, such as B.1.351 (also 501Y.V2) detected in South Africa 6 and P.1 (also 501Y.V3) recently detected in Brazil 7 . We identified a deletion in the ORF1a gene (ORF1a Δ3675-3677) in all three variants, which has not yet been widely detected in other SARS-CoV-2 lineages. Using ORF1a Δ3675-3677 as the primary target and spike Δ69-70 to differentiate, we designed and validated an open source PCR assay to detect SARS-CoV-2 variants of concern 8 . Our assay can be rapidly deployed in laboratories around the world to enhance surveillance for the local emergence spread of B.1.1.7, B.1.351, and P.1.