Development of a tick-borne encephalitis serodiagnostic ELISA using recombinant Fc-antigen fusion proteins.
Current diagnostic tests for tick-borne encephalitis virus (TBEV) infections require high-biosafety facilities for antigen preparation and can cross-react with other flaviviruses. There is, therefore, a need to develop safe, inexpensive serodiagnostic tools with high specificity and sensitivity. In this study, a recombinant plasmid that expresses the membrane and envelope (E) proteins of TBEV fused to the Fc domain of rabbit IgG was constructed and expressed in mammalian cells. The E-Fc proteins were secreted as soluble homodimers, which retained reactivity with anti-TBEV and anti-rabbit IgG antibodies. The E-Fc proteins were then used to develop an enzyme-linked immunosorbent assay (ELISA) to detect TBEV antibodies in rodent and human sera. Compared with the neutralization test, the ELISA had over 90% sensitivity and specificity. In addition, the assay showed no cross-reactivity with Japanese encephalitis virus antibodies. These findings suggest that the E-Fc ELISA may be a useful tool for TBEV serodiagnosis.
Laboratory of Public Health, Graduate School of Veterinary Medicine, Hokkaido University, Japan; Clinical Studies Department, School of Veterinary Medicine, University of Zambia, Zambia.
Nozyechi N
NN
Chidumayo
Laboratory of Public Health, Graduate School of Veterinary Medicine, Hokkaido University, Japan. Electronic address: kyoshii@vetmed.hokuda.ac.jp.
Kentaro
K
Yoshii
Department of Disease Control, School of Veterinary Medicine, University of Zambia, Zambia.
Ngonda
N
Saasa
Laboratory of Public Health, Graduate School of Veterinary Medicine, Hokkaido University, Japan.
Misuki
M
Sakai
Laboratory of Public Health, Graduate School of Veterinary Medicine, Hokkaido University, Japan.
Hiroaki
H
Kariwa
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D000956:Antigens, Viral; D003955:Diagnostic Tests, Routine / Q000379:methods*; D004675:Encephalitis, Tick-Borne / Q000175:diagnosis*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006801:Humans; D007159:Immunologic Tests / Q000379:methods*; D011993:Recombinant Fusion Proteins; D012680:Sensitivity and Specificity
D000914:Antibodies, Viral; D000956:Antigens, Viral; D011993:Recombinant Fusion Proteins
Diagnosis; ELISA; Fc fusion protein; Tick-borne encephalitis virus