title: Survey of Canadian retail pork chops and pork livers for detection of hepatitis E virus, norovirus, and rotavirus using real time RT-PCR.
abstract: Over the past 15 years, hepatitis E virus (HEV), norovirus (NoV), and rotavirus (RV) have been hypothesized to be potentially zoonotic; swine and pork have been suggested as possible human infection sources for all 3 viruses. Our objective was to estimate HEV, NoV, and RV prevalence and load on Canadian retail pork chops and livers. Using the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS) sampling platform, pork livers (n=283) and chops (n=599) were collected, processed, and assayed for the 3 viruses by four collaborating federal laboratories using validated real time reverse transcriptase polymerase chain reactions (qRT-PCR). Follow-up qRT-PCR estimating viral load in genomic copies/g was followed by nested classical RT-PCR and isolate sequencing of a partial segment of the ORF2 gene. Local alignments were performed using MUSCLE (Multiple Sequence Comparison by Log-Expectation); a phylogenetic tree was created. Twenty-five livers and 6 chops were classified 'positive' (thresholds for viral RNA detected in both replicates of the assay) or 'suspect' (thresholds detected in one of two replicates) for HEV. Follow-up qRT-PCR detected HEV on 16 livers, 0 chops, and nested classical RT-PCR, on 14 livers and 0 chops. Initial qRT-PCR classified 12 chops 'suspect' for NoV. Follow-up qRT-PCR detected viral RNA on only one sample with thresholds greater than 40 in both replicates. No amplicon was yielded, and therefore no isolate was sequenced from this sample. Partial ORF2 genes from 14 HEV isolates were sequenced, and compared via sequence identity and phylogenetic analysis with selected human case isolates listed in NCBI-GenBank. Overall, HEV prevalence on retail pork was comparable with other published reports.
authors.affiliation: University of Guelph, P.O. Box 3339, Vermilion, AB, Canada. Electronic address: bwilhelm@uoguelph.ca.
authors.forename: Barbara
authors.initials: B
authors.lastname: Wilhelm
authors.affiliation: Agriculture and Agri-Food Canada, Food Research and Development Centre, 3600 Casavant Blvd. West, St-Hyacinthe, Québec J2S 8E3, Canada.
authors.forename: Danielle
authors.initials: D
authors.lastname: Leblanc
authors.affiliation: Agriculture and Agri-Food Canada, Food Research and Development Centre, 3600 Casavant Blvd. West, St-Hyacinthe, Québec J2S 8E3, Canada.
authors.forename: Alain
authors.initials: A
authors.lastname: Houde
authors.affiliation: Agriculture and Agri-Food Canada, Food Research and Development Centre, 3600 Casavant Blvd. West, St-Hyacinthe, Québec J2S 8E3, Canada.
authors.forename: Julie
authors.initials: J
authors.lastname: Brassard
authors.affiliation: Agriculture and Agri-Food Canada, Food Research and Development Centre, 3600 Casavant Blvd. West, St-Hyacinthe, Québec J2S 8E3, Canada.
authors.forename: Marie-Josée
authors.initials: MJ
authors.lastname: Gagné
authors.affiliation: Health Canada, Health Products & Foods: Québec Region, 1001 St Laurent West, Longueuil, Québec J4K 1C7, Canada.
authors.forename: Daniel
authors.initials: D
authors.lastname: Plante
authors.affiliation: Health Canada, Health Products & Foods: Québec Region, 1001 St Laurent West, Longueuil, Québec J4K 1C7, Canada.
authors.forename: Pascale
authors.initials: P
authors.lastname: Bellon-Gagnon
authors.affiliation: Agriculture and Agri-Food Canada, Lacombe Research Centre, 6000 C & E Trail, Lacombe, Alberta T4L 1W1, Canada.
authors.forename: Tineke H
authors.initials: TH
authors.lastname: Jones
authors.affiliation: Agriculture and Agri-Food Canada, Lacombe Research Centre, 6000 C & E Trail, Lacombe, Alberta T4L 1W1, Canada.
authors.forename: Victoria
authors.initials: V
authors.lastname: Muehlhauser
authors.affiliation: Laboratory for Foodborne Zoonoses, Public Health Agency of Canada, 160 Research Lane, Suite 103, Guelph, Ontario N1G 5B2, Canada; University of Guelph, Ontario Veterinary College, Guelph, Ontario N1G 2W1, Canada.
authors.forename: Nicol
authors.initials: N
authors.lastname: Janecko
authors.affiliation: Laboratory for Foodborne Zoonoses, Public Health Agency of Canada, 160 Research Lane, Suite 103, Guelph, Ontario N1G 5B2, Canada.
authors.forename: Brent
authors.initials: B
authors.lastname: Avery
authors.affiliation: University of Guelph, Ontario Veterinary College, Guelph, Ontario N1G 2W1, Canada.
authors.forename: Andrijana
authors.initials: A
authors.lastname: Rajić
authors.affiliation: University of Guelph, Ontario Veterinary College, Guelph, Ontario N1G 2W1, Canada.
authors.forename: Scott A
authors.initials: SA
authors.lastname: McEwen
mesh_terms: D000818:Animals; D002170:Canada; D005516:Food Microbiology; D016752:Hepatitis E virus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D008460:Meat / Q000821:virology*; D029322:Norovirus / Q000235:genetics / Q000302:isolation & purification*; D010802:Phylogeny; D012367:RNA, Viral / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012401:Rotavirus / Q000235:genetics / Q000302:isolation & purification*; D013552:Swine; D019562:Viral Load; D014764:Viral Proteins / Q000235:genetics
chemical_list: C400971:ORF2 protein, Hepatitis E virus; D012367:RNA, Viral; D014764:Viral Proteins
keywords: Hepatitis E virus; Norovirus; Pork chops; Pork liver; Retail; Rotavirus