Performance of real-time reverse transcription polymerase chain reaction for the detection of foot-and-mouth disease virus during field outbreaks in the United Kingdom in 2007.
Rapid and accurate diagnosis is essential for effective control of foot-and-mouth disease (FMD). The present report describes the practical steps undertaken to deploy a real-time reverse transcription polymerase chain reaction (real-time RT-PCR) to process the samples received during the outbreaks of FMD in the United Kingdom in 2007. Two independent real-time RT-PCR assays targeting different regions (5'UTR and 3D) of the FMD virus (FMDV) genome were used to confirm the presence of FMDV in clinical samples collected from the first infected premises. Once the FMDV strain responsible had been sequenced, a single real-time RT-PCR assay (3D) was selected to test a total of 3,216 samples, including material from all 8 infected premises. Using a 96-well automated system to prepare nucleic acid template, up to 84 samples could be processed within 5 hr of submission, and up to 269 samples were tested per working day. A conservative cut-off was used to designate positive samples, giving rise to an assay specificity of 99.9% or 100% for negative control material or samples collected from negative premises, respectively. For the first time, real-time RT-PCR results were used to recognize preclinical FMD in a cattle herd. Furthermore, during the later stages of the outbreaks, the real-time RT-PCR assay supported an active surveillance program within high-risk cattle herds. To the authors' knowledge, this is the first documented use of real-time RT-PCR as a principal laboratory diagnostic tool following introduction of FMD into a country that was FMD-free (without vaccination) and highlights the advantages of this assay to support control decisions during disease outbreaks.
Reid
Scott M
SM
Institute for Animal Health, Pirbright, Woking, Surrey, United Kingdom. scott.reid@bbsrc.ac.uk
Ebert
Katja
K
Bachanek-Bankowska
Katarzyna
K
Batten
Carrie
C
Sanders
Anna
A
Wright
Caroline
C
Shaw
Andrew E
AE
Ryan
Eoin D
ED
Hutchings
Geoffrey H
GH
Ferris
Nigel P
NP
Paton
David J
DJ
King
Donald P
DP
D000818:Animals; D002417:Cattle; D004196:Disease Outbreaks / Q000662:veterinary*; D005536:Foot-and-Mouth Disease / Q000175:diagnosis / Q000453:epidemiology*; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012680:Sensitivity and Specificity; D006113:United Kingdom / Q000453:epidemiology