Detecting SARS-CoV-2 variants with SNP genotyping

Darlix, Jean-Luc EPH, Harper, Helen, Burridge, Amanda, Winfield, Mark, Finn, Adam, Davidson, Andrew, Matthews, David, Hutchings, Stephanie, Vipond, Barry, Jain, Nisha, The COVID-19 Genomics UK (COG-UK) Consortium, , Allan, John, Bashton, Matthew, Loh, Joshua, Nelson, Andrew, Smith, Darren L., Yew, Wen Chyin, Young, Gregory R., Edwards, Keith and Barker, Gary (2021) Detecting SARS-CoV-2 variants with SNP genotyping. PLoS ONE, 16 (2). e0243185. ISSN 1932-6203

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Abstract

Tracking genetic variations from positive SARS-CoV-2 samples yields crucial information about the number of variants circulating in an outbreak and the possible lines of transmission but sequencing every positive SARS-CoV-2 sample would be prohibitively costly for population-scale test and trace operations. Genotyping is a rapid, high-throughput and low-cost alternative for screening positive SARS-CoV-2 samples in many settings. We have designed a SNP identification pipeline to identify genetic variation using sequenced SARSCoV-2 samples. Our pipeline identifies a minimal marker panel that can define distinct genotypes. To evaluate the system, we developed a genotyping panel to detect variants-identified from SARS-CoV-2 sequences surveyed between March and May 2020 and tested this on 50 stored qRT-PCR positive SARS-CoV-2 clinical samples that had been collected across the South West of the UK in April 2020. The 50 samples split into 15 distinct genotypes and there was a 61.9% probability that any two randomly chosen samples from our set of 50 would have a distinct genotype. In a high throughput laboratory, qRT-PCR positive samples pooled into 384-well plates could be screened with a marker panel at a cost of < £1.50 per sample. Our results demonstrate the usefulness of a SNP genotyping panel to provide a rapid, cost-effective, and reliable way to monitor SARS-CoV-2 variants circulating in an outbreak. Our analysis pipeline is publicly available and will allow for marker panels to be updated periodically as viral genotypes arise or disappear from circulation.

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