Summary
Reverse transcription followed by the polymerase chain reaction method (PCR) allowed the detection of foot-and-mouth disease virus (FMDV), regardless of the serotype. A primer set corresponding to highly conserved regions of the 2B sequence was selected. By combining in a single reaction tube specific primer pairs for FMDV, swine vesicular disease virus (SVDV), encephalomyocarditis virus (EMCV) and bovine viral diarrhea virus (BVDV), all four viruses could be identified and differentiated in one amplification reaction, based on the different lengths of the respective amplified segments. In a similar way, FMDV types O, A, C, SAT 2 and Asia 1 could be identified and differentiated, using primers selected from the 1D (VP1) genome region. All results were confirmed by direct sequencing of the PCR product. The very fast RNA extraction, reverse transcription and PCR permitted us to read the agarose gels within three hours after samples (cell culture isolates as well as clinical material) arrived, which is of great importance in case of an FMDV suspicion. Furthermore, a very high sensitivity was achieved (less than one PFU). Therefore, our powerful detection assay by means of PCR for FMDV as well as for SVDV, EMCV and BVDV, has great advantages compared to the presently used procedures.
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References
O.I.E. Informations sanitaires (1993) 6, p 31
O.I.E. Informations sanitaires (1993) 6, p 74
Bull. OIE (1994) 106, 7, p 494
Brocchi E, Berlinzani A, Gamba D, De Simone F (1995) Development of two novel monoclonal antibody-based ELISAs for the detection of antibodies and the identification of swine isotypes against swine vesicular disease virus. J Virol Methods 52: 155–167
Koenen F, De Clercq K, Lefebvre J, Strobbe R (1994) Reproductive failure in sows following experimental infection with a Belgian EMCV isolate. Vet Microbiol 39: 111–116
Hedger R, Mann J (1989) Swine vesicular disease virus. In: Pensaert M (ed) Virus infections of porcines. Elsevier, Amsterdam Oxford New York Tokyo, pp 214–250
Mann J, Sellers R (1989) Foot-and-mouth disease virus. In: Pensaert M (ed) Virus infections of porcines. Elsevier, Amsterdam Oxford New York Tokyo, pp 259–264
Mann J, Sellers R (1990) Foot-and-mouth disease virus. In: Horzinek M (ed) Virus infections of vertebrates. Elsevier, Amsterdam Oxford New York Tokyo, pp 503–512 (Virus infections of ruminants, vol 3)
Report of the session of the research group of the standing technical committee of the European commission for the control of foot-and-mouth disease (1994), Vienna, pp 8–9
Waldman O (1928). OIE Bull 1: 518–533
Frenkel HS (1947) La culture du virus de la fièvre aphteuse sue l'épithelium de la langue de bovidés. OIE Bull 28: 155–162
Locher F, Suryanarayana V, Tratschin J-D (1995) Rapid detection and characterization of foot-and-mouth disease virus by restriction enzyme and nucleotide sequence analysis of PCR products. J Clin Microbiol 33: 440–444
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, pp 7.01–7.87
Farrell RE (1993) RNA methodologies: a laboratory guide for isolation and characterization vol 13. Academic Press, San Diego
Mayer R, Brown C, House C, House J, Molitor T (1991) Rapid and sensitive detection of foot-and-mouth disease virus in tissues by enzymatic RNA amplification of the polymerase gene. J Virol Methods 34: 161–172
Laor O, Torgersen H, Yadin H, Becker Y (1992) Detection of FMDV RNA amplified by the polymerase chain reaction (PCR). J Virol Methods 36: 197–208
Amaral-Doel CMF, Owen NE, Ferris NP, Kitching RP, Doel TR (1993) Detection of foot-and-mouth disease viral sequences in clinical specimens and ethyleneimine-inactivated preparations by the polymerase chain reaction. Vaccine 11: 415–421
Höfner M, Carpenter W, Donaldson A (1993) Detection of foot-and-mouth disease virus RNA in clinical samples and cell culture isolates by amplification of the capsid coding region. J Virol Methods 42: 53–62
Donn A, Martin LA, Donaldson AI (1994) Improved detection of persistent foot-and-mouth disease infection in cattle by the polymerase chain reaction. J Virol Methods 49: 179–186
Murphy M, Meyer R, Mebus C, Schudel A, Rodriguez M (1994) Analysis of sites of foot-and-mouth disease virus persistence in carrier cattle via the polymerase chain reaction. Arch Virol 136: 299–307
Rodríguez A, Núñez J, Nolasco G, Ponz F, Sobrino F, Sobrino F, de Blas C (1994) Direct PCR detection of foot-and-mouth disease virus. J Virol Methods 47: 345–349
Dinter Z, Morein B (1990)Picornaviridae. In: Horzinek M (ed) Virus infections of vertebrates. Elsevier Amsterdam Oxford New York Tokyo, p 501 (Virus infections of ruminants, vol 3)
Laor O, Yadin H, Dalia C, Becker Y (1991) Detection of foot-and-mouth disease virus in diagnosic material using the PCR method on viral genomic poly-A RNA isolated with oligo dT on magnetic beads. Isr J Vet Med 46: 127–133
Marquardt O, Straub OC, Ah1 R, Haas B (1994) Type-independent detection of FMDV genomes by RT-PCR: practical aspects. Report of the session of the research group of the standing technical committee of the European commission for the control of foot-and-mouth disease, Vienna
Forss S, Strebel K, Beck E, Schaller H (1984) Nucleotide sequence and genome organization of foot-and-mouth disease virus. Nucleic Acids Res 12: 6587–6601
Seechurn P, Knowles N, McCauley J (1990) The complete nucleotide sequence of a pathogenic swine vesicular disease virus. Virus Res 16: 255–274
Palmenberg A, Kirby E, Janda M, Drake N, Duke G, Potratz K, Collett M (1984) The nucleotide and deduced amino acid sequences of the encephalomyocarditis viral poly-protein coding region. Nucleic Acids Res 12: 2969–2985
Collett M, Larson R, Gold C, Strick D, Anderson D, Purchio A (1988) Molecular cloning and nucleotide sequence of the pestivirus bovine viral diarrhea virus. Virology 165: 191–199
Hertig C, Pauli U, Zanoni R, Peterhans E (1991) Detection of bovine viral diarrhea (BVD) virus using the polymerase chain reaction. Vet Microbiol 26: 65–76
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Vangrysperre, W., De Clercq, K. Rapid and sensitive polymerase chain reaction based detection and typing of foot-and-mouth disease virus in clinical samples and cell culture isolates, combined with a simultaneous differentiation with other genomically and/or symptomatically related viruses. Archives of Virology 141, 331–344 (1996). https://doi.org/10.1007/BF01718403
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DOI: https://doi.org/10.1007/BF01718403