Comparison of different RT-qPCR assays for the detection of human and bovine group A rotaviruses and characterization by sequences analysis of genes encoding VP4 and VP7 capsid proteins

J Appl Microbiol. 2013 May;114(5):1435-48. doi: 10.1111/jam.12165. Epub 2013 Mar 1.

Abstract

Aims: The aim of this study was to compare the performance of four RT-qPCR assays for the detection of human and bovine group A rotaviruses and to characterize the positive samples by sequence analysis of VP4 and VP7 genes.

Methods and results: RNA extracted from eight human rotavirus strains, and a panel of 33 human and 25 bovine faecal samples was subjected to different RT-qPCR detection systems. Among these assays, only RT-qPCR primers and probe systems B and C were able to detect all human rotavirus strains from cell culture solutions and faecal samples. However, the results showed that the system C was generally more sensitive by one or two logs than the other RT-qPCR assays tested. With the bovine faecal samples, the most efficient RT-qPCR systems were B and A with the detection in 100 and 92% of samples tested, respectively. Human group A rotavirus G1P[8] and bovine G6P[11] were the most frequently used strains identified in this study. A G3P[9] strain, closely related to a feline rotavirus isolated in the USA, was also discovered in a human rotavirus infection.

Conclusion: The RT-qPCR system B was the only TaqMan assay evaluated in this study able to detect rotavirus RNA in all positive human and bovine faecal samples.

Significance and impact of the study: Utilization of only one RT-qPCR for the detection of human and bovine group A rotaviruses and the possibility of human infection by a feline rotavirus strain.

Publication types

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

MeSH terms

  • Animals
  • Antigens, Viral / genetics*
  • Capsid Proteins / genetics*
  • Cattle
  • Child
  • DNA Primers
  • Feces / virology
  • Genotype
  • Humans
  • Phylogeny
  • RNA, Viral / isolation & purification
  • Reverse Transcriptase Polymerase Chain Reaction / methods*
  • Rotavirus / classification
  • Rotavirus / genetics
  • Rotavirus / isolation & purification*
  • Rotavirus Infections / diagnosis
  • Rotavirus Infections / veterinary
  • Rotavirus Infections / virology
  • Sensitivity and Specificity
  • Sequence Analysis, DNA

Substances

  • Antigens, Viral
  • Capsid Proteins
  • DNA Primers
  • RNA, Viral
  • VP4 protein, Rotavirus
  • VP7 protein, Rotavirus