title: Development of a real-time reverse transcription polymerase chain reaction assay for detection of marine caliciviruses (genus Vesivirus).
abstract: Marine caliciviruses form a distinct lineage within the genus Vesivirus (family Caliciviridae). This group includes vesicular exanthema of swine virus (VESV) and San Miguel sea lion virus (SMSV) and other related viruses which have been proposed to be marine in origin isolated from a variety of terrestrial and marine animals. Rapid and reliable detection of marine caliciviruses is important as these viruses appear to be widespread and can cause vesicular disease in a wide variety of susceptible hosts including pigs and experimentally infected cattle where clinical signs cannot be easily distinguished from foot-and-mouth disease (FMD), swine vesicular disease (SVD) and vesicular stomatitis (VS). A real-time RT-PCR assay targeting conserved nucleotide sequences in the RNA-dependent RNA polymerase (3D) region of the genome successfully detected cell culture-grown virus preparations of more than thirty marine calicivirus serotypes. Only the atypical SMSV serotypes 8 and 12 failed to be detected, which provided further indication of genetic divergence between these and the other calicivirus serotypes said to be marine in origin. The real-time RT-PCR assay also specifically amplified RNA from samples collected following experimental inoculation of pigs with VESV. No cross-reactivity was demonstrated when the assay was tested with RNA prepared from representative viruses of FMD, SVD and VS. The real-time RT-PCR assay described is a sensitive and specific tool for detection and differential diagnosis of these viruses from other vesicular-disease causing viruses.
authors.affiliation: Institute for Animal Health, Pirbright Laboratory, Ash Road, Pirbright, Woking, Surrey GU24 0NF, United Kingdom. scott.reid.@bbsrc.ac.uk
authors.forename: Scott M
authors.initials: SM
authors.lastname: Reid
authors.forename: Donald P
authors.initials: DP
authors.lastname: King
authors.forename: Andrew E
authors.initials: AE
authors.lastname: Shaw
authors.forename: Nick J
authors.initials: NJ
authors.lastname: Knowles
authors.forename: Geoffrey H
authors.initials: GH
authors.lastname: Hutchings
authors.forename: Emily J
authors.initials: EJ
authors.lastname: Cooper
authors.forename: Alvin W
authors.initials: AW
authors.lastname: Smith
authors.forename: Nigel P
authors.initials: NP
authors.lastname: Ferris
mesh_terms: D000818:Animals; D001483:Base Sequence; D002139:Caliciviridae / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D002417:Cattle; D002460:Cell Line; D017931:DNA Primers; D005536:Foot-and-Mouth Disease / Q000235:genetics; D016679:Genome, Viral; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification; D012324:RNA-Dependent RNA Polymerase / Q000737:chemistry; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D046070:Sea Lions; D012680:Sensitivity and Specificity; D012689:Sequence Homology, Nucleic Acid; D012703:Serotyping; D013552:Swine; D013555:Swine Vesicular Disease / Q000235:genetics; D013997:Time Factors; D014720:Vesicular Exanthema of Swine / Q000235:genetics; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics
chemical_list: D017931:DNA Primers; D012367:RNA, Viral; D012324:RNA-Dependent RNA Polymerase