title: The use of African horse sickness virus NS3 protein, expressed in bacteria, as a marker to differentiate infected from vaccinated horses.
abstract: Segment 10 of the double-stranded RNA (dsRNA) genome from African horse sickness virus serotype 4 (AHSV-4) was cloned and sequenced. The sequence of the coding region showed a total length of 667 bp. Nucleotide comparisons showed a 95% sequence similarity between serotypes 4 and 9, and 76% between serotypes 4 and 3. cDNA clones containing the coding region were cloned in the vector pET3xb and expressed in Escherichia coli. The NS3 gene product was synthesised at very high level as an insoluble fusion protein. The recombinant protein was used in a differential ELISA to distinguish horses that were infected with AHSV-4 or vaccinated with live-modified virus from those vaccinated with a purified inactivated vaccine. The results obtained indicate that recombinant NS3 can indeed differentiate between infected and vaccinated animals implying that this recombinant could be developed as a diagnostic reagent, and it would allow the mobility of vaccinated horses. Thus, economical losses associated with this disease could be avoided.
authors.affiliation: Centro de Investigación en Sanidad Animal, CISA-INIA, Valdeolmos, Madrid, Spain.
authors.forename: M D
authors.initials: MD
authors.lastname: Laviada
authors.forename: P
authors.initials: P
authors.lastname: Roy
authors.forename: J M
authors.initials: JM
authors.lastname: Sánchez-Vizcaíno
authors.forename: J I
authors.initials: JI
authors.lastname: Casal
mesh_terms: D000355:African Horse Sickness / Q000175:diagnosis / Q000276:immunology* / Q000821:virology; D000356:African Horse Sickness Virus / Q000235:genetics* / Q000276:immunology*; D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood / Q000276:immunology; D001483:Base Sequence; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D004279:DNA, Viral; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D012367:RNA, Viral; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D017386:Sequence Homology, Amino Acid; D014611:Vaccination / Q000662:veterinary; D014709:Vero Cells; D017361:Viral Nonstructural Proteins / Q000235:genetics* / Q000276:immunology
chemical_list: D000914:Antibodies, Viral; D004279:DNA, Viral; D012367:RNA, Viral; D011993:Recombinant Fusion Proteins; D017361:Viral Nonstructural Proteins