A Portable Reverse Transcription Recombinase Polymerase Amplification Assay for Rapid Detection of Foot-and-Mouth Disease Virus
Open Access
- 20 August 2013
- journal article
- research article
- Published by Public Library of Science (PLoS) in PLOS ONE
- Vol. 8 (8), e71642
- https://doi.org/10.1371/journal.pone.0071642
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.This publication has 27 references indexed in Scilit:
- Rapid Detection of HIV-1 Proviral DNA for Early Infant Diagnosis Using Recombinase Polymerase AmplificationmBio, 2013
- Emergence of Foot-and-Mouth Disease Virus SAT 2 in Egypt During 2012Transboundary and Emerging Diseases, 2012
- Rapid typing of foot-and-mouth disease serotype Asia 1 by reverse transcription loop-mediated isothermal amplificationVirology Journal, 2011
- Development and laboratory validation of a lateral flow device for the detection of foot-and-mouth disease virus in clinical samplesJournal of Virological Methods, 2009
- Foot-and-Mouth Disease Virus Serotype A in EgyptEmerging Infectious Diseases, 2007
- Novel reverse transcription loop-mediated isothermal amplification for rapid detection of foot-and-mouth disease virusArchiv für die gesamte Virusforschung, 2006
- Sequence variation can affect the performance of minor groove binder TaqMan probes in viral diagnostic assaysJournal of Clinical Virology, 2006
- Molecular epidemiology of foot-and-mouth disease virusVirus Research, 2003
- Differentiation of the seven serotypes of foot-and-mouth disease virus by reverse transcriptase polymerase chain reactionJournal of Virological Methods, 1997
- Detection of specific polymerase chain reaction product by utilizing the 5'----3' exonuclease activity of Thermus aquaticus DNA polymerase.Proceedings of the National Academy of Sciences of the United States of America, 1991