title: Highly specific confirmatory western blot test for African swine fever virus antibody detection using the recombinant virus protein p54.
abstract: A Western blot technique using a recombinant protein has been developed to confirm positive results obtained in African swine fever (ASF)-specific antibody detection by ELISA. The new confirmatory Western blot is based on the use of protein p54, one of the most antigenic ASF virus structural proteins, expressed in Escherichia coli fused to the N-terminus of MS2 polymerase. The recombinant Western blot assay was highly specific and equally sensitive for ASF virus-infected pigs detection as the conventional Western blot, which uses virus-induced proteins ranging in molecular weight between 23 and 35 kDa. The novel Western blot assay provides a simpler interpretation of the test, eliminates the possibility of false-positive reactions produced by cellular compounds that contaminate the antigen employed in the conventional technique, and avoids the use of live virus in antigen production.
authors.affiliation: Centro de Investigación en Sanidad Animal (CISA-INIA), Valdeolmos, Madrid, Spain.
authors.forename: C
authors.initials: C
authors.lastname: Alcaraz
authors.forename: F
authors.initials: F
authors.lastname: Rodriguez
authors.forename: J M
authors.initials: JM
authors.lastname: Oviedo
authors.forename: A
authors.initials: A
authors.lastname: Eiras
authors.forename: M
authors.initials: M
authors.lastname: De Diego
authors.forename: C
authors.initials: C
authors.lastname: Alonso
authors.forename: J M
authors.initials: JM
authors.lastname: Escribano
mesh_terms: D000357:African Swine Fever / Q000175:diagnosis*; D000358:African Swine Fever Virus; D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis*; D001483:Base Sequence; D015153:Blotting, Western / Q000379:methods; D002460:Cell Line; D003001:Cloning, Molecular; D017931:DNA Primers; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D000882:Haplorhini; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000276:immunology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology; D012680:Sensitivity and Specificity; D013552:Swine; D015678:Viral Structural Proteins / Q000096:biosynthesis / Q000276:immunology*
chemical_list: D000914:Antibodies, Viral; D017931:DNA Primers; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D015678:Viral Structural Proteins; C093921:p54 protein, African swine fever virus