title: 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.
abstract: OBJECTIVE
  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.
  
  
  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.
  
  
  CONCLUSIONS
  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.
  
  
  CONCLUSIONS
  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.
authors: Agriculture and Agri-Food Canada, Food Research and Development Centre, Saint-Hyacinthe, QC, Canada. Pierre.Ward@agr.gc.ca
authors: P
authors: P
authors: Ward
authors: E
authors: E
authors: Poitras
authors: D
authors: D
authors: Leblanc
authors: C A
authors: CA
authors: Gagnon
authors: J
authors: J
authors: Brassard
authors: A
authors: A
authors: Houde
mesh_terms: D000818:Animals; D000956:Antigens, Viral / Q000235:genetics*; D036022:Capsid Proteins / Q000235:genetics*; D002417:Cattle; D002648:Child; D017931:DNA Primers; D005243:Feces / Q000821:virology; D005838:Genotype; D006801:Humans; D010802:Phylogeny; D012367:RNA, Viral / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012401:Rotavirus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D012400:Rotavirus Infections / Q000175:diagnosis / Q000662:veterinary / Q000821:virology; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA
chemical_list: D000956:Antigens, Viral; D036022:Capsid Proteins; D017931:DNA Primers; D012367:RNA, Viral; C060339:VP4 protein, Rotavirus; C050779:VP7 protein, Rotavirus