title: Development of a serodiagnostic multi-species ELISA against tick-borne encephalitis virus using subviral particles.
abstract: Tick-borne encephalitis virus (TBEV) is a zoonotic agent causing severe encephalitis in humans. A wide range of animal species could be infected with TBEV in endemic areas. A serological survey of wild animals is effective in identifying TBEV-endemic areas. Safe, simple, and reliable TBEV serodiagnostic tools are needed to test animals. In this study, ELISA was developed to detect anti-TBEV specific antibodies in multi-species of animals, using recombinant subviral particles (SPs) with an affinity tag and protein A/G. A Strep-tag was fused at the N terminus of the E protein of the plasmid coding TBEV prME. The E proteins with Strep-tag were secreted as SPs, of which Strep-tag was exposed on the surface. The tagged E proteins were associated with prM. The SPs with Strep-tag were applied as the antigen of ELISA, and TBEV-specific antibodies were detected by the protein A/G. Compared to neutralization test results, the ELISA showed 96.8% sensitivity and 97.7% specificity in rodents and 95.1% sensitivity and 96.0% specificity in humans, without cross-reactivity with antibodies to Japanese encephalitis virus. These results indicate that our ELISA would be useful to detect TBE-specific antibodies in a wide range of animal species.
authors.affiliation: Laboratory of Public Health, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Hokkaido 060-0818, Japan.
authors.forename: Eri
authors.initials: E
authors.lastname: Inagaki
authors.affiliation: Laboratory of Public Health, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Hokkaido 060-0818, Japan.
authors.forename: Mizuki
authors.initials: M
authors.lastname: Sakai
authors.affiliation: Laboratory of Public Health, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Hokkaido 060-0818, Japan.
authors.forename: Minato
authors.initials: M
authors.lastname: Hirano
authors.affiliation: Laboratory of Public Health, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Hokkaido 060-0818, Japan.
authors.forename: Memi
authors.initials: M
authors.lastname: Muto
authors.affiliation: Laboratory of Public Health, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Hokkaido 060-0818, Japan.
authors.forename: Shintaro
authors.initials: S
authors.lastname: Kobayashi
authors.affiliation: Laboratory of Public Health, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Hokkaido 060-0818, Japan.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Kariwa
authors.affiliation: Laboratory of Public Health, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Hokkaido 060-0818, Japan. Electronic address: kyoshii@vetmed.hokudai.ac.jp.
authors.forename: Kentaro
authors.initials: K
authors.lastname: Yoshii
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D004669:Encephalitis Viruses, Tick-Borne / Q000276:immunology*; D004675:Encephalitis, Tick-Borne / Q000175:diagnosis* / Q000662:veterinary*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D012377:Rodentia; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000379:methods*
chemical_list: D000914:Antibodies, Viral
keywords: ELISA; Flavivirus; Subviral particles; Tick-borne encephalitis virus