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
Article first published online: 1 MAR 2013
© Her Majesty the Queen in Right of Canada . Reproduced with the permission of the Minister of Agriculture and Agri-food Canada.
Journal of Applied Microbiology
Volume 114, Issue 5, pages 1435–1448, May 2013
How to Cite
Ward, P., Poitras, E., Leblanc, D., Gagnon, C.A., Brassard, J. and Houde, A. (2013), 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. Journal of Applied Microbiology, 114: 1435–1448. doi: 10.1111/jam.12165
- Issue published online: 15 APR 2013
- Article first published online: 1 MAR 2013
- Accepted manuscript online: 19 FEB 2013 12:49PM EST
- Manuscript Accepted: 23 JAN 2013
- Manuscript Revised: 11 JAN 2013
- Manuscript Received: 18 OCT 2012
- Agriculture and Agri-Food Canada Research Branch. Grant Number: RPBI # 1485
- capsid protein sequences;
- group A rotaviruses;
- phylogenetic analysis;
- RT-qPCR assays
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 and bovine G6P were the most frequently used strains identified in this study. A G3P strain, closely related to a feline rotavirus isolated in the USA, was also discovered in a human rotavirus infection.
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.