title: Pan-serotypic detection of foot-and-mouth disease virus using a minor groove binder probe reverse transcription polymerase chain reaction assay.
abstract: A novel assay for the pan-serotypic detection of foot-and-mouth disease virus (FMDV) was designed using a 5' conjugated minor groove binder (MGB) probe real-time RT-PCR system. This assay targets the 3D region of the FMDV genome and is capable of detecting 20 copies of a transcribed RNA standard. The linear range of the test was eight logs from 2 × 10¹ to 2 × 10⁸ copies and amplification time was approximately 2 h. Using a panel of 83 RNA samples from representative FMDV isolates, the diagnostic sensitivity of this test was shown to be equivalent to a TaqMan real-time RT-PCR that targets the 5' untranslated region of FMDV. Furthermore, the assay does not detect viruses causing similar clinical diseases in pigs such as swine vesicular disease virus and vesicular stomatitis virus, nor does it detect marine caliciviruses causing vesicular exanthema. The development of this assay provides a useful tool for the differential diagnosis of FMD, potentially for use in statutory or emergency testing programmes, or for detection of FMDV RNA in research applications.
authors.affiliation: Veterinary Sciences Division, Agri-Food and Biosciences Institute, Stormont, Belfast BT4 3SD, UK. john.mckillen@afbini.gov.uk
authors.forename: John
authors.initials: J
authors.lastname: McKillen
authors.forename: Michael
authors.initials: M
authors.lastname: McMenamy
authors.forename: Scott M
authors.initials: SM
authors.lastname: Reid
authors.forename: Catherine
authors.initials: C
authors.lastname: Duffy
authors.forename: Bernt
authors.initials: B
authors.lastname: Hjertner
authors.forename: Donald P
authors.initials: DP
authors.lastname: King
authors.forename: Sándor
authors.initials: S
authors.lastname: Bélak
authors.forename: Michael
authors.initials: M
authors.lastname: Welsh
authors.forename: Gordon
authors.initials: G
authors.lastname: Allan
mesh_terms: D000818:Animals; D019411:Clinical Laboratory Techniques / Q000379:methods*; D005536:Foot-and-Mouth Disease / Q000175:diagnosis* / Q000821:virology; D029544:Foot-and-Mouth Disease Virus / Q000235:genetics / Q000302:isolation & purification*; D015345:Oligonucleotide Probes / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014773:Virology / Q000379:methods*
chemical_list: D015345:Oligonucleotide Probes