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Clinical and Diagnostic Laboratory Immunology, November 2004, p. 1120-1129, Vol. 11, No. 6
1071-412X/04/$08.00+0     DOI: 10.1128/CDLI.11.6.1120-1129.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Serological Method Using Recombinant S2 Protein To Differentiate Equine Infectious Anemia Virus (EIAV)-Infected and EIAV-Vaccinated Horses

Sha Jin,1 Charles J. Issel,2 and Ronald C. Montelaro1*

Department of Molecular Genetics and Biochemistry, School of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania,1 Department of Veterinary Science, Gluck Equine Research Center, University of Kentucky, Lexington, Kentucky2

Received 25 June 2004/ Returned for modification 2 September 2004/ Accepted 9 September 2004

We recently reported a highly protective attenuated live virus vaccine for equine infectious anemia virus (EIAV) based on a proviral construct (EIAVUK{Delta}S2) with a genetically engineered mutation in the viral S2 gene that eliminates expression of this accessory protein. While the EIAVUK{Delta}S2 vaccine provides protection from detectable infection by experimental challenge with highly virulent virus, the potential for commercial application of this vaccine is complicated by the fact that horses inoculated with the EIAVUK{Delta}S2 vaccine strain become seropositive in various reference diagnostic assays based on detection of antibodies to virion core or envelope proteins. To address this issue, we describe here the development and optimization of a new serologic EIAV diagnostic enzyme-linked immunosorbent assay (ELISA) to detect serum antibodies to the EIAV S2 protein that are produced in infected horses but not in horses inoculated with the EIAVUK{Delta}S2 vaccine virus. The test S2 protein antigen was developed using the S2 gene sequence from the EIAVUK strain of virus and a series of modifications to facilitate production and purification of the diagnostic antigen, designated HS2G. Using this HS2G as antigen, we describe the development of an affinity ELISA that provides a sensitive and specific detection of S2-specific serum antibodies in experimentally and field-infected horses (22 of 24), without detectable reactivity with immune serum from uninfected (12 of 12) or vaccinated (29 of 29) horses. These data indicate that the S2-based diagnostic ELISA has the potential to accurately differentiate horses infected with EIAV from horses inoculated with an attenuated EIAV vaccine strain with a mutant S2 gene.

* Corresponding author. Mailing address: W1144 Biomedical Science Tower, Department of Molecular Genetics and Biochemistry, School of Medicine, University of Pittsburgh, Pittsburgh, PA 15261. Phone: (412) 648-8869. Fax: (412) 383-8859. E-mail: rmont{at}pitt.edu.

Clinical and Diagnostic Laboratory Immunology, November 2004, p. 1120-1129, Vol. 11, No. 6
1071-412X/04/$08.00+0     DOI: 10.1128/CDLI.11.6.1120-1129.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Jin, S., Chen, C., Montelaro, R. C. (2005). Equine Infectious Anemia Virus Gag p9 Function in Early Steps of Virus Infection and Provirus Production. J. Virol. 79: 8793-8801 [Abstract] [Full Text]  


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