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Journal of Clinical Microbiology, September 2004, p. 4101-4110, Vol. 42, No. 9
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.9.4101-4110.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

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

Michael R. Holbrook,^* Robert E. Shope, and Alan D. T. Barrett

Department of Pathology, and Center for Biodefense and Emerging Infectious Diseases, University of Texas Medical Branch, Galveston Texas

Received 20 August 2003/ Accepted 20 May 2004

The serological diagnosis of infection by flaviviruses is complicated by the presence of flavivirus cross-reactive 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.

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* Corresponding author. Present address: Department of Pathology, 1 University of New Mexico, MSC 08 4640, Albuquerque, NM 87131. Phone: (505) 272-5872. Fax: (505) 272-9912. E-mail: MHolbrook{at}salud.unm.edu.

Deceased.

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Journal of Clinical Microbiology, September 2004, p. 4101-4110, Vol. 42, No. 9
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.9.4101-4110.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

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