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PCR assay to enhance global surveillance for SARS-CoV-2 variants of concern

Chantal B.F. Vogels, Mallery Breban, Tara Alpert, Mary E. Petrone, Anne E. Watkins, Emma B. Hodcroft, Christopher E. Mason, Gaurav Khullar, Jessica Metti, Joel T. Dudley, Matthew J. MacKay, Megan Nash, Jianhui Wang, Chen Liu, Pei Hui, Steven Murphy, Caleb Neal, Eva Laszlo, Marie L. Landry, Anthony Muyombwe, Randy Downing, Jafar Razeq, Richard A. Neher, Joseph R. Fauver, Nathan D. Grubaugh
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With the emergence of SARS-CoV-2 variants that may increase transmissibility and/or cause escape from immune responses1–3, 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 UK4,5 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 Africa6 and P.1 (also 501Y.V3) recently detected in Brazil7. 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 concern8. 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.

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Publicado en el sitio 2021-02-06 10:13:46

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