Use of Recombinant E Protein Domain III-Based Enzyme-Linked Immunosorbent Assays for Differentiation of Tick-Borne Encephalitis Serocomplex Flaviviruses from Mosquito-Borne Flaviviruses

Abstract

The serological diagnosis of infection by flaviviruses is complicated by the presence of flavivirus crossreactive antibodies that produce false-positive results for flavivirus infections, especially in regions where more than one virus is endemic. Current diagnostic reagents for tick-borne flavivirus infection have been found to cross-react with yellow fever-or dengue virus-positive sera. This study utilized recombinant flavivirus E protein domain 3 (rE-D3) as a diagnostic reagent to differentiate between infection by mosquito-and tick-borne flaviviruses. This study found that the use of rE-D3 in an enzyme-linked immunosorbent assay (ELISA)-based format allowed the differentiation between serum specific for either mosquito-or tick-borne flaviviruses, but not among the members of the tick-borne encephalitis (TBE) serocomplex of flaviviruses. Sera derived against several TBE serocomplex rE-D3 were found to cross-react with heterologous rE-D3 within the TBE serocomplex, but not with those from mosquito-borne flaviviruses, in both Western blots and ELISAs. Mouse hyperimmune sera generated against TBE serocomplex viruses were also found to react specifically with TBE serocomplex rE-D3, but not with rE-D3 from mosquito-borne viruses and vice versa. When a similar test using virus-derived antigen was performed, a loss of both specificity and sensitivity was observed. These results indicate that flavivirus rE-D3 would be a useful reagent for the detection of infection by TBE serocomplex flaviviruses, several of which are potential biothreat agents, but would not provide the ability to differentiate among infections by separate members of the serocomplex.

Genetic Determinants of Sindbis Virus Mosquito Infection Are Associated with a Highly Conserved Alphavirus and Flavivirus Envelope Sequence

“…Because the Mut-5-like alphavirus and flavivirus residues align spatially but their sequences are not well conserved (Fig. 4) and even encompass a deletion region that corresponds solely to tick-borne flaviviruses (14), this region may be hypothesized to constitute a variable region (VR) involved in mosquito infections and is termed VR-I in Fig. 4 (SINV E2 aa 116 to 122, DENV-2 E aa 382 to 388).…”

Tick-Borne Encephalitis Virus, Kyrgyzstan

“…We analyzed 302 rodents and insectivores for immunoglobulin (Ig) G and IgM to TBEV by using recombinant antigen of domain III from the envelope (E) protein of Kumlinge and Powassan viruses (6). This assay is specifi c for the tick-borne fl avivirus group and lacks cross-reactivity that occurs with other assays (7).…”

Antibody persistence after two vaccinations with either FSME-IMMUN® Junior or ENCEPUR® Children followed by third vaccination with FSME-IMMUN® Junior

“…The antigenic components of both inactivated vaccines are highly homologous with only a few variable amino acid positions in the envelope (E) protein gene which do not affect functionally important sequences. 8,9 According to the manufacturers' recommendations, the conventional pediatric primary vaccination schedule consists of 3 doses of the vaccine: the first and second vaccinations administered one to three months apart, followed by the third vaccination 5-12 mo later for FSME-IMMUN ® Junior and 9-12 mo later for Encepur ® Children. A rapid immunization schedule is available for both vaccines; with FSME-IMMUN ® Junior, the first two vaccinations are www.landesbioscience.com Human Vaccines & Immunotherapeutics 737 administered 14 d apart, followed by the third vaccination 5-12 mo later.…”