Enzyme-linked immunosorbent assay using recombinant antigens expressed in mammalian cells for serodiagnosis of tick-borne encephalitis.
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.
Yoshii
Kentarou
K
Department of Environmental Veterinary Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060-0818, Japan.
Hayasaka
Daisuke
D
Goto
Akiko
A
Obara
Mayumi
M
Araki
Koichi
K
Yoshimatsu
Kumiko
K
Arikawa
Jiro
J
Ivanov
Leonoid
L
Mizutani
Tetsuya
T
Kariwa
Hiroaki
H
Takashima
Ikuo
I
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
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