A portable reverse transcription recombinase polymerase amplification assay for rapid detection of foot-and-mouth disease virus

PLoS One. 2013 Aug 20;8(8):e71642. doi: 10.1371/journal.pone.0071642. eCollection 2013.

Abstract

Foot-and-mouth disease (FMD) is a trans-boundary viral disease of livestock, which causes huge economic losses and constitutes a serious infectious threat for livestock farming worldwide. Early diagnosis of FMD helps to diminish its impact by adequate outbreak management. In this study, we describe the development of a real-time reverse transcription recombinase polymerase amplification (RT-RPA) assay for the detection of FMD virus (FMDV). The FMDV RT-RPA design targeted the 3D gene of FMDV and a 260 nt molecular RNA standard was used for assay validation. The RT-RPA assay was fast (4-10 minutes) and the analytical sensitivity was determined at 1436 RNA molecules detected by probit regression analysis. The FMDV RT-RPA assay detected RNA prepared from all seven FMDV serotypes but did not detect classical swine fever virus or swine vesicular disease virus. The FMDV RT-RPA assay was used in the field during the recent FMD outbreak in Egypt. In clinical samples, reverse transcription polymerase chain reaction (RT-PCR) and RT-RPA showed a diagnostic sensitivity of 100% and 98%, respectively. In conclusion, FMDV RT-RPA was quicker and much easier to handle in the field than real-time RT-PCR. Thus RT-RPA could be easily implemented to perform diagnostics at quarantine stations or farms for rapid spot-of-infection detection.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Computer Systems
  • DNA Primers / metabolism
  • Disease Outbreaks
  • Egypt / epidemiology
  • Foot-and-Mouth Disease / diagnosis*
  • Foot-and-Mouth Disease / epidemiology
  • Foot-and-Mouth Disease / virology*
  • Foot-and-Mouth Disease Virus / genetics*
  • Foot-and-Mouth Disease Virus / isolation & purification*
  • Recombinases / metabolism*
  • Reverse Transcriptase Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity

Substances

  • DNA Primers
  • Recombinases

Grants and funding

The study was funded by Alexander von Humboldt Foundation, Bonn, Germany (project ID: 3.4 - Fokoop-DEU/1144102). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.