title: Typing of viral hemorrhagic septicemia virus by monoclonal antibodies.
abstract: Seven mAbs with specific reaction patterns against each of the four genotypes and eight subtypes of viral hemorrhagic septicemia virus (VHSV) were produced, aiming to establish an immunoassay for typing VHSV isolates according to their genotype. Among the mAbs, VHS-1.24 reacted with all genotypes except genotype Ie, whilst mAb VHS-9.23 reacted with all genotypes except genotype III. mAb VHS-3.80 reacted with genotypes Ib, Ic, Id and II. mAb VHS-7.57 reacted with genotypes II and IVa, and mAb VHS-5.18 with genotype Ib only. Interestingly, mAb VHS-3.75 reacted with all of the genotype III isolates except a rainbow trout-pathogenic isolate from the west coast of Norway, and reacted in addition with the IVb isolate, CA-NB00-01, from the east coast of the USA. Finally, mAb VHS-1.88 reacted with all genotype IVb isolates from the Great Lakes, but not with CA-NB00-01. In conclusion, we can distinguish between all four genotypes and between five of eight subtypes of VHSV by testing isolates in immunoassay using a panel of nine mAbs. By Western blotting and transfection of cell cultures, it was shown that mAb VHS-1.24 recognized an epitope on the viral phosphoprotein (P), whilst all others recognized antigenic determinants on the nucleoprotein (N). From amino acid alignments of the various genotypes and subtypes of VHSV isolates, it was possible to determine the epitope specificity of mAb VHS-1.24 to be aa 32-34 in the P-protein; the specificities of mAbs VHS-3.80, VHS-7.57 and VHS-3.75 were found to be aa 43 and 45-48, aa 117 and 121, and aa 103, 118 and 121 of the N-protein, respectively.
authors.affiliation: Tamaki Station, Aquatic Animal Health Division, National Research Institute of Aquaculture, Fisheries Research Agency, 224-1 Hiruta, Tamaki, Mie 519-0423, Japan.
authors.forename: Takafumi
authors.initials: T
authors.lastname: Ito
authors.affiliation: Tamaki Station, Aquatic Animal Health Division, National Research Institute of Aquaculture, Fisheries Research Agency, 224-1 Hiruta, Tamaki, Mie 519-0423, Japan.
authors.forename: Jun
authors.initials: J
authors.lastname: Kurita
authors.affiliation: Aquatic Animal Health Division, National Research Institute of Aquaculture, Fisheries Research Agency, Minami-Ise, Mie 516-0193, Japan.
authors.forename: Motohiko
authors.initials: M
authors.lastname: Sano
authors.affiliation: National Veterinary Institute, Technical University of Denmark, Hangøvej 2, DK-8200 Århus N, Denmark.
authors.forename: Helle Frank
authors.initials: HF
authors.lastname: Skall
authors.affiliation: National Veterinary Institute, Technical University of Denmark, Hangøvej 2, DK-8200 Århus N, Denmark.
authors.forename: Niels
authors.initials: N
authors.lastname: Lorenzen
authors.affiliation: National Veterinary Institute, Technical University of Denmark, Hangøvej 2, DK-8200 Århus N, Denmark.
authors.forename: Katja
authors.initials: K
authors.lastname: Einer-Jensen
authors.affiliation: National Veterinary Institute, Technical University of Denmark, Hangøvej 2, DK-8200 Århus N, Denmark.
authors.forename: Niels Jørgen
authors.initials: NJ
authors.lastname: Olesen
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D058846:Antibodies, Monoclonal, Murine-Derived / Q000276:immunology*; D000918:Antibody Specificity; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000235:genetics / Q000276:immunology; D019084:Fluorescent Antibody Technique, Indirect; D005838:Genotype; D006825:Hybridomas / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D029141:Novirhabdovirus / Q000145:classification* / Q000235:genetics / Q000276:immunology* / Q000472:pathogenicity; D017386:Sequence Homology, Amino Acid; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology
chemical_list: D058846:Antibodies, Monoclonal, Murine-Derived; D000939:Epitopes; D014764:Viral Proteins