ABSTRACT
The complete nucleotide sequence of foot-and-mouth disease virus (FMDV) O/FRA/1/2001 (bovine isolate) was determined from five cDNA clones covering most of the genome and compared with the British porcine isolate (O/UKG/35/2001) it originated from. Seven substitutions, out of which three resulted in amino acid changes (in the leader protease, 3A protein and 3D RNA-dependent RNA polymerase sequences) were identified and confirmed by direct sequencing of RT-PCR products obtained from in vitro infected cells and skin vesicles of an infected cow. RACE amplification allowed determination of the exact 3' end of the genome. These changes and possibly the unusual 3A substitution between the British and its French derivate may account for the consecutive species shifts.
Subject(s)
Foot-and-Mouth Disease Virus/genetics , Genome, Viral , Amino Acid Sequence , Amino Acid Substitution , Animals , Antigens, Viral/genetics , Base Sequence , Cattle , Cattle Diseases/virology , DNA, Complementary/chemistry , DNA, Complementary/isolation & purification , DNA, Viral/chemistry , DNA, Viral/isolation & purification , Endopeptidases/genetics , Foot-and-Mouth Disease/virology , Foot-and-Mouth Disease Virus/isolation & purification , France , Molecular Sequence Data , Mutation, Missense , Point Mutation , United Kingdom , Viral Nonstructural Proteins/geneticsABSTRACT
Foot-and-mouth disease virus (FMDV) is an important veterinary pathogen which can cause widespread epidemics. Due to the high antigenic variability of FMDV, it is important to undertake mutation analysis under immunological pressure. To study the bovine antibody response at a molecular level, phage display technology was used to produce bovine anti-FMDV Fabs. CH1-VH chains with FMDV specific binding could be isolated after selection from a library made from vaccinated cattle. Though their involvement in the bovine immune response remains to be ascertained, it is planned to express the five different selected VH domains in bacterial or insect systems as sequence homologies with integrin beta6 chain could shed light on the basis of FMDV type receptor specificities.
Subject(s)
Antibodies, Viral/immunology , Cattle Diseases/virology , Foot-and-Mouth Disease Virus/immunology , Foot-and-Mouth Disease/immunology , Immunoglobulin Fragments/isolation & purification , Immunoglobulin Variable Region/isolation & purification , Amino Acid Sequence , Animals , Antibodies, Viral/genetics , Base Sequence , Blotting, Western/veterinary , Cattle , Cattle Diseases/immunology , Cloning, Molecular , DNA, Viral/chemistry , DNA, Viral/genetics , Enzyme-Linked Immunosorbent Assay/veterinary , Epitopes/immunology , Foot-and-Mouth Disease Virus/genetics , Gene Library , Immunoglobulin Fragments/genetics , Immunoglobulin Fragments/immunology , Immunoglobulin Variable Region/genetics , Immunoglobulin Variable Region/immunology , Molecular Sequence Data , Peptide Library , Polymerase Chain Reaction/veterinaryABSTRACT
Foot-and-mouth disease (FMD) diagnostic methods are reviewed. As the presence of clinical signs alone is inconclusive, laboratory diagnosis should always be carried out. The presence of FMD virus can be demonstrated by cell culture isolation, complement fixation test, ELISA or the more recent polymerase chain reaction (PCR) method. Serological diagnosis is also a valuable tool. The virus neutralization test has been replaced by ELISA and the antibody response to some viral non-structural proteins allows to discriminate between vaccinated and infected animals on a herd basis. More rapid and accurate tests as well as an earlier detection system in preclinical state are still needed.
