title: A portable reverse transcription recombinase polymerase amplification assay for rapid detection of foot-and-mouth disease virus.
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.
authors: Department of Virology, University Medical Center, Goettingen, Germany ; Virology Department, Faculty of Veterinary Medicine, Mansoura University, Mansoura, Egypt.
authors: Ahmed
authors: A
authors: Abd El Wahed
authors: Ayman
authors: A
authors: El-Deeb
authors: Mohamed
authors: M
authors: El-Tholoth
authors: Hanaa
authors: H
authors: Abd El Kader
authors: Abeer
authors: A
authors: Ahmed
authors: Sayed
authors: S
authors: Hassan
authors: Bernd
authors: B
authors: Hoffmann
authors: Bernd
authors: B
authors: Haas
authors: Mohamed A
authors: MA
authors: Shalaby
authors: Frank T
authors: FT
authors: Hufert
authors: Manfred
authors: M
authors: Weidmann
mesh_terms: D000818:Animals; D003199:Computer Systems; D017931:DNA Primers / Q000378:metabolism; D004196:Disease Outbreaks; D004534:Egypt / Q000453:epidemiology; D005536:Foot-and-Mouth Disease / Q000175:diagnosis* / Q000453:epidemiology / Q000821:virology*; D029544:Foot-and-Mouth Disease Virus / Q000235:genetics* / Q000302:isolation & purification*; D045522:Recombinases / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D017931:DNA Primers; D045522:Recombinases
references: Virus Res. 2003 Jan;91(1):65-80
references: 12527438
references: Transbound Emerg Dis. 2012 Dec;59(6):476-81
references: 23025522
references: Virol J. 2011 Oct 31;8:489
references: 22040459
references: J Virol Methods. 1997 Aug;67(1):35-44
references: 9274816
references: Clin Microbiol Rev. 2004 Apr;17(2):465-93
references: 15084510
references: N Z Vet J. 2002 Apr;50(2):46-55
references: 16032210
references: J Vet Diagn Invest. 2007 Jan;19(1):9-20
references: 17459827
references: J Virol Methods. 1999 Dec;83(1-2):113-23
references: 10598089
references: Proc Natl Acad Sci U S A. 1991 Aug 15;88(16):7276-80
references: 1871133
references: J Clin Virol. 2012 Aug;54(4):308-12
references: 22683006
references: J Clin Microbiol. 2003 Apr;41(4):1565-8
references: 12682146
references: Methods Mol Biol. 2007;402:3-34
references: 17951788
references: J Clin Microbiol. 2012 Jul;50(7):2234-8
references: 22518861
references: J Am Vet Med Assoc. 2002 Jun 1;220(11):1636-42
references: 12051502
references: J Virol Methods. 2009 Jan;155(1):10-7
references: 18848845
references: Expert Rev Vaccines. 2011 Mar;10(3):377-87
references: 21434805
references: Nature. 1981 Apr 30;290(5809):800-2
references: 6261157
references: Virol J. 2011 Nov 09;8:510
references: 22070774
references: mBio. 2013 Apr 02;4(2):
references: 23549916
references: Vet Rec. 2011 Sep 24;169(13):336
references: 21846685
references: Transbound Emerg Dis. 2010 Apr;57(1-2):19-21
references: 20537095
references: Arch Virol. 2006 Jun;151(6):1093-106
references: 16453084
references: Vet Med Int. 2011;2011:905768
references: 21776357
references: PLoS Biol. 2006 Jul;4(7):e204
references: 16756388
references: J Clin Virol. 2006 Jan;35(1):81-3
references: 16002331
references: J Virol. 2012 Aug;86(16):8901-2
references: 22843860
references: Emerg Infect Dis. 2007 Oct;13(10):1593-6
references: 18258017