| 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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MedLine Citation:
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PMID: 12588687 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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. |
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Authors:
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Margarita Sáiz; Diana B De La Morena; Esther Blanco; José I Núñez; Rufino Fernández; José M Sánchez-Vizcaíno |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Veterinary research Volume: 34 ISSN: 0928-4249 ISO Abbreviation: Vet. Res. Publication Date: 2003 Jan-Feb |
Date Detail:
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Created Date: 2003-02-17 Completed Date: 2003-05-23 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 9309551 Medline TA: Vet Res Country: France |
Other Details:
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Languages: eng Pagination: 105-17 Citation Subset: IM |
Affiliation:
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Centro de Investigación en Sanidad Animal, INIA, 28130 Valdeolmos, Madrid, Spain. saiz@inia.es |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Base Sequence Foot-and-Mouth Disease / diagnosis*, virology* Foot-and-Mouth Disease Virus / genetics*, isolation & purification* Genes, Viral / genetics Phylogeny Restriction Mapping / methods, veterinary* Reverse Transcriptase Polymerase Chain Reaction / methods, veterinary* Sensitivity and Specificity Sequence Alignment Sequence Analysis, DNA / veterinary* Swine / virology |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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