Detection of all seven serotypes of foot-and-mouth disease virus by real-time, fluorogenic reverse transcription polymerase chain reaction assay.
A fluorogenic RT-PCR (5'-nuclease probe-based) assay using a primer/probe set designed from the internal ribosomal entry site region of the virus genome was developed for the specific detection of all seven serotypes of foot-and-mouth disease (FMD) virus in epithelial suspensions and cell culture virus preparations. The reverse transcription polymerase chain reaction (RT-PCR) specifically detected FMD virus in sample submissions from the UK 2001 FMD outbreak with greater sensitivity than our conventional RT-PCR procedure and our routine diagnostic procedures of ELISA and virus isolation in cell culture. The fluorogenic RT-PCR provides relatively fast results, enables a quantitative assessment to be made of virus amounts and can handle more samples and/or replicates of samples in a single assay than the conventional RT-PCR procedure. Therefore it is seen as a valuable tool to complement the routine diagnostic procedures for FMD virus diagnosis.
Reid
Scott M
SM
Institute for Animal Health, Pirbright Laboratory, Ash Road, Pirbright, Woking, GU24 0NF, Surrey, UK.
Ferris
Nigel P
NP
Hutchings
Geoffrey H
GH
Zhang
Zhidong
Z
Belsham
Graham J
GJ
Alexandersen
Soren
S
D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D015342:DNA Probes; D004196:Disease Outbreaks; D004797:Enzyme-Linked Immunosorbent Assay; D004847:Epithelial Cells; D005456:Fluorescent Dyes; D005536:Foot-and-Mouth Disease / Q000175:diagnosis* / Q000453:epidemiology / Q000821:virology; D029544:Foot-and-Mouth Disease Virus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D012367:RNA, Viral / Q000032:analysis; D012015:Reference Standards; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012703:Serotyping; D006113:United Kingdom / Q000453:epidemiology; D014776:Virus Cultivation
D015342:DNA Probes; D005456:Fluorescent Dyes; D012367:RNA, Viral