Investigating the biological and technical origins of unknown bases in the S region of the SARS-CoV-2 Delta variant genome sequences

Borcard, Loïc; Gempeler, Sonja; Terrazos Miani, Miguel A.; Baumann, Christian; Grädel, Carole; Dijkman, Ronald; Suter-Riniker, Franziska; Leib, Stephen L; Bittel, Pascal; Neuenschwander, Stefan; Ramette, Alban (2021). Investigating the biological and technical origins of unknown bases in the S region of the SARS-CoV-2 Delta variant genome sequences (medRxiv). Cold Spring Harbor Laboratory 10.1101/2021.09.09.21262951

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We are reporting on the observation of a large, under-sequenced region of the S gene of the SARS-CoV2 Delta variant genomes, identified in sequences originating from various sequencing centres worldwide (e.g. USA, India, England, Switzerland, France, Germany). This poorly sequenced region was identified from the early phases of the Delta variant spread and the phenomenon is still ongoing. As many commonly-used protocols rely on amplicon-based sequencing procedures, we investigated the likely origin of the issue. We established its biological origin as resulting from mutations in the viral genomes at primer binding sites. We designed and evaluated new PCR primers to circumvent this issue in order to complement the ARTIC v3 set, and validated their performance for the sequencing of circulating Delta variants.

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