title: Development of a novel real-time RT-PCR assay to detect Seneca Valley virus-1 associated with emerging cases of vesicular disease in pigs.
abstract: Seneca Valley virus 1 (SVV-1) can cause vesicular disease that is clinically indistinguishable from foot-and-mouth disease, vesicular stomatitis and swine vesicular disease. SVV-1-associated disease has been identified in pigs in several countries, namely USA, Canada, Brazil and China. Diagnostic tests are required to reliably detect this emerging virus, and this report describes the development and evaluation of a novel real-time (r) reverse-transcription (RT) PCR assay (rRT-PCR), targeting the viral polymerase gene (3D) of SVV-1. This new assay detected all historical and contemporary SVV-1 isolates examined (n=8), while no cross-reactivity was observed with nucleic acid templates prepared from other vesicular disease viruses or common swine pathogens. The analytical sensitivity of the rRT-PCR was 0.79 TCID50/ml and the limit of detection was equivalent using two different rRT-PCR master-mixes. The performance of the test was further evaluated using pig nasal (n=25) and rectal swab samples (n=25), where concordant results compared to virus sequencing were generated for 43/50 samples. The availability of this assay, will enable laboratories to rapidly detect SVV-1 in cases of vesicular disease in pigs, negated for notifiable diseases, and could enable existing knowledge gaps to be investigated surrounding the natural epidemiology of SVV-1.
authors.affiliation: The Pirbright Institute, Ash Road, Pirbright, Woking, Surrey GU24 0NF, United Kingdom.
authors.forename: Veronica L
authors.initials: VL
authors.lastname: Fowler
authors.affiliation: Kansas State Veterinary Diagnostic Laboratory, Kansas State University, 1800 Denison Avenue, Manhattan, KS 66506, United States.
authors.forename: Russell H
authors.initials: RH
authors.lastname: Ransburgh
authors.affiliation: Kansas State Veterinary Diagnostic Laboratory, Kansas State University, 1800 Denison Avenue, Manhattan, KS 66506, United States.
authors.forename: Elizabeth G
authors.initials: EG
authors.lastname: Poulsen
authors.affiliation: The Pirbright Institute, Ash Road, Pirbright, Woking, Surrey GU24 0NF, United Kingdom.
authors.forename: Jemma
authors.initials: J
authors.lastname: Wadsworth
authors.affiliation: The Pirbright Institute, Ash Road, Pirbright, Woking, Surrey GU24 0NF, United Kingdom.
authors.forename: Donald P
authors.initials: DP
authors.lastname: King
authors.affiliation: The Pirbright Institute, Ash Road, Pirbright, Woking, Surrey GU24 0NF, United Kingdom.
authors.forename: Valerie
authors.initials: V
authors.lastname: Mioulet
authors.affiliation: The Pirbright Institute, Ash Road, Pirbright, Woking, Surrey GU24 0NF, United Kingdom.
authors.forename: Nick J
authors.initials: NJ
authors.lastname: Knowles
authors.affiliation: Animal and Plant Health Agency, Rougham Hill, Bury St Edmunds, Suffolk IP30 9AN, United Kingdom.
authors.forename: Susanna
authors.initials: S
authors.lastname: Williamson
authors.affiliation: Kansas State Veterinary Diagnostic Laboratory, Kansas State University, 1800 Denison Avenue, Manhattan, KS 66506, United States; Department of Diagnostic Medicine/Pathobiology, Kansas State University, 1800 Denison Avenue, Manhattan, KS 66506, United States.
authors.forename: Xuming
authors.initials: X
authors.lastname: Liu
authors.affiliation: Kansas State Veterinary Diagnostic Laboratory, Kansas State University, 1800 Denison Avenue, Manhattan, KS 66506, United States; Department of Diagnostic Medicine/Pathobiology, Kansas State University, 1800 Denison Avenue, Manhattan, KS 66506, United States.
authors.forename: Gary A
authors.initials: GA
authors.lastname: Anderson
authors.affiliation: Department of Diagnostic Medicine/Pathobiology, Kansas State University, 1800 Denison Avenue, Manhattan, KS 66506, United States.
authors.forename: Ying
authors.initials: Y
authors.lastname: Fang
authors.affiliation: Kansas State Veterinary Diagnostic Laboratory, Kansas State University, 1800 Denison Avenue, Manhattan, KS 66506, United States; Department of Diagnostic Medicine/Pathobiology, Kansas State University, 1800 Denison Avenue, Manhattan, KS 66506, United States. Electronic address: jbai@vet.ksu.edu.
authors.forename: Jianfa
authors.initials: J
authors.lastname: Bai
mesh_terms: D000818:Animals; D021821:Communicable Diseases, Emerging / Q000175:diagnosis / Q000662:veterinary / Q000821:virology; D057230:Limit of Detection; D009666:Nose / Q000821:virology; D010849:Picornaviridae / Q000235:genetics*; D010850:Picornaviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D012367:RNA, Viral / Q000302:isolation & purification; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012007:Rectum / Q000821:virology; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis* / Q000821:virology
chemical_list: D012367:RNA, Viral
keywords: Real-time; Seneca Valley virus; Senecavirus A; Swine; Veterinary diagnostics; rRT-PCR