A universal genome sequencing method for rotavirus A from human fecal samples which identifies segment reassortment and multi-genotype mixed infection

BMC Genomics. 2017 Apr 24;18(1):324. doi: 10.1186/s12864-017-3714-6.

Abstract

Background: Genomic characterization of rotavirus (RoV) has not been adopted at large-scale due to the complexity of obtaining sequences for all 11 segments, particularly when feces are used as starting material.

Methods: To overcome these limitations, we developed a novel RoV capture and genome sequencing method combining commercial enzyme immunoassay plates and a set of routinely used reagents.

Results: Our approach had a 100% success rate, producing >90% genome coverage for diverse RoV present in fecal samples (Ct < 30).

Conclusions: This method provides a novel, reproducible and comparatively simple approach for genomic RoV characterization and could be scaled-up for use in global RoV surveillance systems.

Trial registration (prospectively registered): Current Controlled Trials ISRCTN88101063 . Date of registration: 14/06/2012.

Keywords: Antibody capture; Co-infection; Genome sequencing; Genomics; Phylogenetics; Reassortment; Rotavirus A.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA, Complementary / genetics
  • Feces / virology*
  • Genome, Viral / genetics
  • Genomics / methods*
  • Genotype*
  • Humans
  • Phylogeny
  • Reassortant Viruses / genetics*
  • Reassortant Viruses / physiology
  • Rotavirus / genetics*
  • Rotavirus / physiology
  • Sequence Analysis, RNA / methods*
  • Viral Load

Substances

  • DNA, Complementary

Associated data

  • ISRCTN/ISRCTN88101063