Author:
Andersen P.,Barksdale S.,Barclay R.A.,Smith N.,Fernandes J.,Besse K.,Goldfarb D.,Barbero R.,Dunlap R.,Jones-Roe T.,Kelly R.,Miao S.,Ruhunusiri C.,Munns A.,Mosavi S.,Sanson L.,Munns D.,Sahoo S.,Swahn O.,Hull K.,White D.,Kolb K.,Noroozi F.,Seelam J.,Patnaik A.,Lepene B.
Abstract
AbstractPresented 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.
Funder
Centers for Disease Control and Prevention
Publisher
Springer Science and Business Media LLC
Reference35 articles.
1. WHO Coronavirus (COVID-19) Dashboard. https://covid19.who.int.
2. Greninger AL et al. (2021) Clinical and infection prevention applications of SARS-CoV-2 genotyping an IDSA/ASM consensus review document. J Clin Microbiol, 0, 01659-21.
3. Robishaw, J. D. et al. Genomic surveillance to combat COVID-19: Challenges and opportunities. Lancet Microbe 2, e481–e484 (2021).
4. Vankadari, N. Overwhelming mutations or SNPs of SARS-CoV-2: A point of caution. Gene 752, 144792 (2020).
5. Safdar, N., Moreno, G. K., Braun, K. M., Friedrich, T. C. & O’Connor, D. H. Using virus sequencing to determine source of SARS-CoV-2 transmission for healthcare worker. Emerg. Infect. Dis. 26, 2489–2491 (2020).
Cited by
4 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献