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Journal of Veterinary Diagnostic Investigation, Vol 18, Issue 1, 93-97
Copyright © 2006 by American Association of Veterinary Laboratory Diagnosticians

Articles

Detection of foot-and-mouth disease virus: comparative diagnostic sensitivity of two independent real-time reverse transcription-polymerase chain reaction assays

DP King, NP Ferris, AE Shaw, SM Reid, GH Hutchings, AC Giuffre, JM Robida, JD Callahan, WM Nelson, and TR Beckham

Institute for Animal Health, Pirbright Laboratory, Surrey, UK. donald.king@bbsrc.ac.uk

Rapid and accurate diagnosis is central to the effective control of foot-and-mouth disease (FMD). It is now recognized that reverse-transcription polymerase chain reaction (RT-PCR) assays can play an important role in the routine detection of FMD virus (FMDV) in clinical samples. The aim of this study was to compare the ability of 2 independent real-time RT-PCR (rRT-PCR) assays targeting the 5' untranslated region (5'UTR) and RNA polymerase (3D) to detect FMDV in clinical samples. There was concordance between the results generated by the 2 assays for 88.1% (347 of 394) of RNA samples extracted from suspensions of epithelial tissue obtained from suspect FMD cases. The comparison between the 2 tests highlighted 19 FMDV isolates (13 for the 5'UTR and 6 for the 3D assay), which failed to produce a signal in 1 assay but gave a positive signal in the other. The sequence of the genomic targets of selected isolates highlighted nucleotide substitutions in the primer or probe regions, thereby providing an explanation for negative results generated in the rRT-PCR assays. These data illustrate the importance of the continuous monitoring of circulating FMDV field strains to ensure the design of the rRT-PCR assay remains fit for purpose and suggest that the use of multiple diagnostic targets could further enhance the sensitivity of molecular methods for the detection of FMDV.


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B. J. Hindson, S. M. Reid, B. R. Baker, K. Ebert, N. P. Ferris, L. F. B. Tammero, R. J. Lenhoff, P. Naraghi-Arani, E. A. Vitalis, T. R. Slezak, et al.
Diagnostic Evaluation of Multiplexed Reverse Transcription-PCR Microsphere Array Assay for Detection of Foot-and-Mouth and Look-Alike Disease Viruses
J. Clin. Microbiol., March 1, 2008; 46(3): 1081 - 1089.
[Abstract] [Full Text] [PDF]


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