title: Development of a minor groove binder assay for real-time one-step RT-PCR detection of swine vesicular disease virus.
abstract: The design and development of a 5' conjugated minor groove binder (MGB) probe real-time RT-PCR assay are described for rapid, sensitive and specific detection of swine vesicular disease virus (SVDV) RNA. The assay is designed to target the 2C gene of the SVDV genome and is capable of detecting 2×10(2) copies of an RNA standard per reaction. It does not detect any of the other RNA viruses that cause vesicular disease in pigs, or the human enterovirus, Coxsackie B5 virus (CVB5) which is closely related antigenically to SVDV. The linear range of this test was from 2×10(2) to 2×10(8) copies/μl. The assay is rapid and can detect SVDV RNA in just over 3.5 h including the time required for nucleic acid extraction. The development of this assay provides a useful tool for the differential diagnosis of SVD or for the detection of SVDV in research applications. This study demonstrates the suitability of MGB probes as a real-time PCR chemistry for the diagnosis of swine vesicular disease.
authors.affiliation: School of Biological Sciences, Queens University Belfast, Belfast, Northern Ireland, UK. m.mcmenamy@qub.ac.uk
authors.forename: M J
authors.initials: MJ
authors.lastname: McMenamy
authors.forename: J
authors.initials: J
authors.lastname: McKillen
authors.forename: S M
authors.initials: SM
authors.lastname: Reid
authors.forename: B
authors.initials: B
authors.lastname: Hjertner
authors.forename: D P
authors.initials: DP
authors.lastname: King
authors.forename: B
authors.initials: B
authors.lastname: Adair
authors.forename: G
authors.initials: G
authors.lastname: Allan
mesh_terms: D000818:Animals; D017931:DNA Primers / Q000235:genetics*; D029822:Enterovirus B, Human / Q000235:genetics / Q000302:isolation & purification*; D004769:Enterovirus Infections / Q000175:diagnosis* / Q000821:virology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis* / Q000821:virology*; D013997:Time Factors; D014773:Virology / Q000379:methods*
chemical_list: D017931:DNA Primers