title: Enzyme-linked immunosorbent assay using recombinant antigens expressed in mammalian cells for serodiagnosis of tick-borne encephalitis.
abstract: A recombinant plasmid that expresses the tick-borne encephalitis (TBE) virus premembrane (prM) and envelope (E) proteins in mammalian cells was constructed. Recombinant proteins retained antigenic and conformational structures similar to those of native virus proteins, and transfected cells released virus-like particles (VLPs), which were 1.13-1.14 g/ml in density and 20-30 nm in diameter, into the culture medium. Recombinant E proteins were used for the development of an enzyme-linked immunosorbent assay (ELISA) to detect TBE virus-specific IgM and IgG antibodies in serum. The results of this ELISA correlated well with the results of commercial ELISA, when tested with 95 serum samples from clinically TBE-suspected patients. In addition, ELISA using recombinant antigens showed no cross-reactivity against serum from Japanese encephalitis (JE) patients, despite the cross-reactivity shown by commercial ELISA systems. These observations indicated that this newly developed ELISA system could distinguish tick-borne encephalitis from Japanese encephalitis infection, and that it constitutes a useful and safe alternative to conventional ELISA systems.
authors.affiliation: Department of Environmental Veterinary Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060-0818, Japan.
authors.forename: Kentarou
authors.initials: K
authors.lastname: Yoshii
authors.forename: Daisuke
authors.initials: D
authors.lastname: Hayasaka
authors.forename: Akiko
authors.initials: A
authors.lastname: Goto
authors.forename: Mayumi
authors.initials: M
authors.lastname: Obara
authors.forename: Koichi
authors.initials: K
authors.lastname: Araki
authors.forename: Kumiko
authors.initials: K
authors.lastname: Yoshimatsu
authors.forename: Jiro
authors.initials: J
authors.lastname: Arikawa
authors.forename: Leonoid
authors.initials: L
authors.lastname: Ivanov
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Mizutani
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Kariwa
authors.forename: Ikuo
authors.initials: I
authors.lastname: Takashima
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D000956:Antigens, Viral / Q000235:genetics; D002460:Cell Line; D006224:Cricetinae; D003429:Cross Reactions; D004669:Encephalitis Viruses, Tick-Borne / Q000235:genetics / Q000276:immunology*; D004672:Encephalitis, Japanese / Q000175:diagnosis / Q000276:immunology / Q000821:virology; D004675:Encephalitis, Tick-Borne / Q000175:diagnosis* / Q000276:immunology / Q000821:virology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood; D007075:Immunoglobulin M / Q000097:blood; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics; D012698:Serologic Tests / Q000379:methods; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology; D014773:Virology / Q000379:methods
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C074932:prM protein, Flavivirus; C065696:glycoprotein E, Flavivirus