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Comparison of different RT ‐q PCR assays for the detection of human and bovine group A rotaviruses and characterization by sequences analysis of genes encoding VP 4 and VP 7 capsid proteins
Author(s) -
Ward P.,
Poitras E.,
Leblanc D.,
Gag C.A.,
Brassard J.,
Houde A.
Publication year - 2013
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/jam.12165
Subject(s) - capsid , biology , gene , virology , reoviridae , rotavirus , virus , genetics , microbiology and biotechnology
Aims The aim of this study was to compare the performance of four RT ‐q PCR assays for the detection of human and bovine group A rotaviruses and to characterize the positive samples by sequence analysis of VP 4 and VP 7 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 ‐q PCR detection systems. Among these assays, only RT ‐q PCR 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 ‐q PCR assays tested. With the bovine faecal samples, the most efficient RT ‐q PCR systems were B and A with the detection in 100 and 92% of samples tested, respectively. Human group A rotavirus G 1 P [8] and bovine G 6 P [11] were the most frequently used strains identified in this study. A G 3 P [9] strain, closely related to a feline rotavirus isolated in the USA , was also discovered in a human rotavirus infection. Conclusion The RT ‐q PCR system B was the only T aq M an 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 ‐q PCR for the detection of human and bovine group A rotaviruses and the possibility of human infection by a feline rotavirus strain.