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Archives of Virology

, Volume 151, Issue 6, pp 1093–1106 | Cite as

Novel reverse transcription loop-mediated isothermal amplification for rapid detection of foot-and-mouth disease virus

  • J. P. Dukes
  • D. P. King
  • S. Alexandersen
Article

Summary.

Speed is paramount in the diagnosis of foot-and-mouth disease (FMD) and simplicity is required if a test is to be deployed in the field. The development of a one-step, reverse transcription loop-mediated amplification (RT-LAMP) assay enables FMD virus (FMDV) to be detected in under an hour in a single tube without thermal cycling. A fragment of the 3D RNA polymerase gene of the virus is amplified at 65 °C in the presence of a primer mixture and both reverse transcriptase and Bst DNA polymerase. Compared with real-time RT-PCR, RT-LAMP was consistently faster, and ten copies of FMDV transcript were detected in twenty-two minutes. Amplification products were detected by visual inspection, agarose gel electrophoresis, or in real-time by the addition of a fluorescent dye. The specificity of the reaction was demonstrated by the absence of amplification of RNA from other viruses that cause vesicular diseases and from that of genetically related picornaviruses. Diagnostic sensitivity was validated by the amplification of reference FMDV strains and archival material from field cases of FMD. In comparison with the performance of the established diagnostic TaqMan® assay, RT-LAMP appears to be sensitive, rapid, specific, and cost-effective, with the potential for field deployment and use by developing countries for FMDV surveillance.

Keywords

Reverse Transcription Rapid Detection Diagnostic Sensitivity Single Tube Polymerase Gene 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • J. P. Dukes
    • 1
  • D. P. King
    • 1
  • S. Alexandersen
    • 2
  1. 1.Institute for Animal Health, Pirbright LaboratoryPirbrightU.K.
  2. 2.Department of VirologyDanish Institute for Food and Veterinary ResearchKalvehaveDenmark

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