Antigenic characterization of Hantaan and Seoul virus nucleocapsid proteins expressed by recombinant baculovirus: application of a truncated protein, lacking an antigenic region common to the two viruses, as a serotyping antigen.
Hantaan virus (HTN) and Seoul virus (SEO) are members of the genus Hantavirus in the family Bunyaviridae and are causative agents of hemorrhagic fever with renal syndrome. The complete and truncated nucleocapsid proteins (NP) of HTN and SEO were expressed by a recombinant baculovirus system. Antigenic characterization of the NP using monoclonal antibodies (MAbs) indicated that the binding sites for the serotype-specific MAbs were located between amino acids (aa) 155 and 429. A Western blot assay indicated that the serotype-specific epitopes were conformation dependent. An indirect immunofluorescence antibody (IFA) assay with the truncated NP (aa 155 to 429) was able to distinguish convalescent-phase sera from HTN and SEO patients. However, the antibody titers with the truncated NP were lower than those with the whole NP. The truncated NP of SEO (aa 155 to 429) could be used as an enzyme-linked immunosorbent assay (ELISA) antigen, but the truncated NP from HTN lost its reactivity when used for ELISA. The IFA assay using baculovirus-expressed truncated NP as an antigen is a rapid, simple, and safe test for distinguishing between HTN and SEO infections by serotype.
Hokkaido University School of Medicine, Hokkaido University, Sapporo 060, Japan.
M
M
Morii
K
K
Yoshimatsu
J
J
Arikawa
G
G
Zhou
H
H
Kariwa
I
I
Takashima
D000328:Adult; D000818:Animals; D000911:Antibodies, Monoclonal; D002460:Cell Line; D003001:Cloning, Molecular / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect; D018063:Hantaan virus / Q000145:classification*; D006476:Orthohantavirus / Q000145:classification*; D018778:Hantavirus Infections / Q000097:blood / Q000175:diagnosis*; D006480:Hemorrhagic Fever with Renal Syndrome / Q000097:blood / Q000175:diagnosis*; D006801:Humans; D007313:Insecta; D008297:Male; D008875:Middle Aged; D019590:Nucleocapsid Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology*; D017924:Nucleopolyhedroviruses; D016133:Polymerase Chain Reaction / Q000379:methods; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology; D017384:Sequence Deletion; D012703:Serotyping; D014162:Transfection
D000911:Antibodies, Monoclonal; D000939:Epitopes; D019590:Nucleocapsid Proteins; D011994:Recombinant Proteins