Magnetic hydrogel particles improve nanopore sequencing of SARS-CoV-2 and other respiratory viruses
P. Andersen, Stephanie M. Barksdale, R. Barclay, Natalie Smith, JOSÉ P. V. FERNANDES, K. Besse, Daniel Goldfarb, Robbie Barbero, Ross Dunlap, Tara Jones-Roe, Ross E. A. Kelly, Shida Miao, Chamodya Ruhunusiri, A. R. I. Munns, Sayed Mohammad R. Mosavi, L. Sanson, Denton Munns, Soumya Sibani Sahoo, Olivia Swahn, Katrusha Hull, DAVID O. WHITE, K. Kolb, F. Noroozi, J. Seelam, Anurag Patnaik, Benjamin Lepene
Abstract
Presented here is a magnetic hydrogel particle enabled workflow for capturing and concentrating SARS-CoV-2 from diagnostic remnant swab samples that significantly improves sequencing results using the Oxford Nanopore Technologies MinION sequencing platform. Our approach utilizes a novel affinity-based magnetic hydrogel particle, circumventing low input sample volumes and allowing for both rapid manual and automated high throughput workflows that are compatible with Nanopore sequencing. This approach enhances standard RNA extraction protocols, providing up to 40 × improvements in viral mapped reads, and improves sequencing coverage by 20-80% from lower titer diagnostic remnant samples. Furthermore, we demonstrate that this approach works for contrived influenza virus and respiratory syncytial virus samples, suggesting that it can be used to identify and improve sequencing results of multiple viruses in VTM samples. These methods can be performed manually or on a KingFisher automation platform.