Probe capture enrichment next-generation sequencing of complete foot-and-mouth disease virus genomes in clinical samples
Nagendrakumar B Singanallur, Danielle E Anderson, October M Sessions, Uma S Kamaraj, Timothy R Bowden, Jacquelyn Horsington, Christopher Cowled, Lin-Fa Wang, Wilna Vosloo
Journal of Virological Methods | ELSEVIER | Published : 2019
Next-generation sequencing (NGS) techniques offer an unprecedented "step-change" increase in the quantity and quality of sequence data rapidly generated from a sample and can be applied to obtain ultra-deep coverage of viral genomes. This is not possible with the routinely used Sanger sequencing method that gives the consensus reads, or by cloning approaches. In this study, a targeted-enrichment methodology for the simultaneous acquisition of complete foot-and-mouth disease virus (FMDV) genomes directly from clinical samples is presented. Biotinylated oligonucleotide probes (120 nt) were used to capture and enrich viral RNA following library preparation. To create a virus capture panel targe..View full abstract
Awarded by Australian Government under the FMD Risk Management Project Phase 1
Awarded by Australian Government under the FMD Risk Management Project Phase 2
The authors thank the Duke-NUS Medical School ABSL3 facility staff, especially Benson Ng and Velraj Sivalingam, for their assistance with the importation of FMDV samples. The authors are grateful to Xiao Fang Lim for technical assistance. Funding for this work was provided through the Meat & Livestock Australia Donor Company matching arrangements with the Australian Government under the FMD Risk Management Project Phase 1 (P.PSH.0558) and 2 (P.PSH.0652).