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Detection of foot-and-mouth disease virus from culture and clinical samples by reverse transcription-PCR coupled to restriction enzyme and sequence analysis

Veterinary Research publica este artículo de investigación

1 de enero de 2003

A reverse transcription-PCR (RT-PCR) method is presented for the highly sensitive and specific detection of foot-and-mouth disease virus (FMDV). A primer pair flanking a region of the viral polymerase gene (3D) corresponding to the C-terminus of the protein was designed and a single step RT-PCR reaction was developed. The assay allowed the detection of viral RNA from a variety of animal samples and from a wide range of FMDV isolates of different origins and serotypes. The presence of an Ahd I restriction site within the amplicon in 96% of the isolates analyzed allowed an additional confirmation step of the positive reactions by a simple digestion yielding characteristic fragment sizes. The set of primers described here was suitable for direct sequencing of the PCR product (290 bp), and the nucleotide sequences corresponding to the SAT 1 and SAT 3 strains were determined. The segment amplified, when used in phylogenetic studies, allowed the clustering of SAT isolates and the rest of FMDV strains as two separate lineages




Saiz M., de la Morena DB., Blanco E., Nunez JL., Fernandez R. y Sanchez-Vizcaino JM..




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Detection of foot-and-mouth disease virus from culture and clinical samples by reverse transcription-PCR coupled to restriction enzyme and sequence analysis

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Detection of foot-and-mouth disease virus from culture and clinical samples by reverse transcription-PCR coupled to restriction enzyme and sequence analysis



Participantes:

Universidad ComplutenseDepartamento de Sanidad Animal. Facultad de Veterinaria. Universidad Complutense (UCM).

Universidad ComplutenseServicio de Inmunología Viral y Medicina Preventiva (SUAT). Centro de Vigilancia Sanitaria Veterinaria (VISAVET). Universidad Complutense (UCM).

Instituto Nacional de Investigación y Tecnología Agraria y AlimentariaCentro de Investigación en Sanidad Animal (CISA). Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA).

Consejo Superior de Investigaciones CientíficasCentro de Biología Molecular Severo Ochoa (CBMSO). Consejo Superior de Investigaciones Científicas (CSIC). Universidad Autónoma de Madrid (UAM).







Veterinary Research
FACTOR YEAR Q
1.991 2003

NLMID: 9309551

PMID: 12588687

ISSN: 0928-4249



TÍTULO: Detection of foot-and-mouth disease virus from culture and clinical samples by reverse transcription-PCR coupled to restriction enzyme and sequence analysis


REVISTA: Vet Res


NUMERACIÓN: 34(1):105-17


AÑO: 2003


EDITORIAL: BMC


AUTORES: Saiz M., de la Morena DB., Blanco E., Nunez JL., Fernandez R. and Sanchez-Vizcaino JM..


José Manuel Sánchez-Vizcaíno Rodríguez

DOI: https://doi.org/10.1051/vetres:2002059


CITA ESTA PUBLICACIÓN:

Saiz M., de la Morena DB., Blanco E., Nunez JL., Fernandez R. y Sanchez-Vizcaino JM. Detection of foot-and-mouth disease virus from culture and clinical samples by reverse transcription-PCR coupled to restriction enzyme and sequence analysis. Veterinary Research. 34(1):105-17. 2003. (A). ISSN: 0928-4249. DOI: 10.1051/vetres:2002059


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