title: Antigen-capture ELISA for the detection of Rift Valley fever virus nucleoprotein using new monoclonal antibodies.
abstract: Monoclonal antibodies (MAbs) raised against the nucleoprotein (NP) of Rift Valley fever virus (RVFV) were developed, and an antigen-capture enzyme-linked immunosorbent assay (Ag-capture ELISA) system was developed for the detection of RVFV NP. The assay detected RVFV antigen from culture supernatants containing as little as 7.8-31.3 pfu per 100 μl. Reactivity with various truncated NPs indicated that MAb C10-54 bound only to the full-length NP, probably due to recognition of a conformational epitope, whereas MAbs G2-36 and D5-59 bound to a linear epitope ranging from amino acid residues 195-201 in the C-terminal region. Based on the alignments of the amino acid sequence of RVFV NP, the epitope regions of MAbs G2-36 and D5-59 were completely conserved among all RVFV strains. These results suggest that the MAbs are applicable to the Ag-capture ELISA for the diagnosis of RVFV infections.
authors.affiliation: Department of Virology 1, National Institute of Infectious Diseases, 4-7-1 Gakuen, Musashimurayama, Tokyo 208-0011, Japan.
authors.forename: Shuetsu
authors.initials: S
authors.lastname: Fukushi
authors.forename: Mina
authors.initials: M
authors.lastname: Nakauchi
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Mizutani
authors.forename: Masayuki
authors.initials: M
authors.lastname: Saijo
authors.forename: Ichiro
authors.initials: I
authors.lastname: Kurane
authors.forename: Shigeru
authors.initials: S
authors.lastname: Morikawa
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology; D000914:Antibodies, Viral / Q000737:chemistry / Q000276:immunology; D000956:Antigens, Viral / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D018604:Epitope Mapping; D000939:Epitopes / Q000737:chemistry; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D019590:Nucleocapsid Proteins / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D012295:Rift Valley Fever / Q000175:diagnosis* / Q000276:immunology; D012296:Rift Valley fever virus / Q000276:immunology*; D016415:Sequence Alignment; D018411:Spodoptera; D014709:Vero Cells
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D000939:Epitopes; D019590:Nucleocapsid Proteins