Detection of bovine group a rotavirus using rapid antigen detection kits, rt-PCR and next-generation DNA sequencing

J Vet Med Sci. 2013 Dec 30;75(12):1651-5. doi: 10.1292/jvms.13-0265. Epub 2013 Aug 2.

Abstract

We investigated the sensitivity of human rotavirus rapid antigen detection (RAD) kits, RT-PCR and next-generation DNA sequencing (NGS) for detection of bovine group A rotavirus (RVA). The Dipstick 'Eiken' Rota (Dipstick) showed the highest sensitivity out of the seven RAD kits against all selected strains in limited dilution analyses, which was consistent with the results for equine rotavirus previously reported. RT-PCR had 10⁰-10³-fold higher sensitivity than the Dipstick. NGS using thirteen RT-PCR-negative fecal samples revealed that all samples yielded RVA reads and especially that two of them covered all 11 genome segments. Moreover, mapping reads to reference sequences allowed genotyping. The NGS would be sensitive and useful for analysis of less dependent on specific primers and screening of genotypes.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, Viral / isolation & purification
  • Cattle
  • Cattle Diseases / diagnosis*
  • Cattle Diseases / virology*
  • High-Throughput Nucleotide Sequencing / veterinary
  • Reagent Kits, Diagnostic / veterinary*
  • Reagent Kits, Diagnostic / virology
  • Reverse Transcriptase Polymerase Chain Reaction / veterinary
  • Rotavirus / genetics*
  • Rotavirus / immunology*
  • Rotavirus Infections / diagnosis
  • Rotavirus Infections / veterinary*
  • Sensitivity and Specificity

Substances

  • Antigens, Viral
  • Reagent Kits, Diagnostic