COVseq is a cost-effective workflow for mass-scale SARS-CoV-2 genomic surveillance

Abstract

While mass-scale vaccination campaigns are ongoing worldwide, genomic surveillance of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is critical to monitor the emergence and global spread of viral variants of concern (VOC). Here, we present a streamlined workflow—COVseq—which can be used to generate highly multiplexed sequencing libraries compatible with Illumina platforms from hundreds of SARS-CoV-2 samples in parallel, in a rapid and cost-effective manner. We benchmarked COVseq against a standard library preparation method (NEBNext) on 29 SARS-CoV-2 positive samples, reaching 95.4% of concordance between single-nucleotide variants detected by both methods. Application of COVseq to 245 additional SARS-CoV-2 positive samples demonstrated the ability of the method to reliably detect emergent VOC as well as its compatibility with downstream phylogenetic analyses. A cost analysis showed that COVseq could be used to sequence thousands of samples at less than 15 USD per sample, including library preparation and sequencing costs. We conclude that COVseq is a versatile and scalable method that is immediately applicable for SARS-CoV-2 genomic surveillance and easily adaptable to other pathogens such as influenza viruses.

COVseq is a cost-effective workflow for mass-scale SARS-CoV-2 genomic surveillance

“…1a, b, Supplementary Data 1, 2, and "Methods"). A step-by-step COVseq protocol is available in Supplementary Methods and at Protocol Exchange 22 . We first determined the breadth of coverage of the SARS-CoV-2 genome that could theoretically be achieved using two frequently cutting restriction enzymes compatible with CUTseq 21 .…”

“…COVseq. A detailed step-by-step COVseq protocol is available in the Supplementary Information and at Protocol Exchange 22 . Below, we briefly describe two COVseq workflows, depending on the number of samples to be processed.…”