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Journal of Veterinary Diagnostic Investigation Vol. 20 Issue 6, 789-791
Copyright © 2008 by the American Association of Veterinary Laboratory Diagnosticians
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Brief Research Reports

Evaluation of an indirect enzyme-linked immunosorbent assay for routine screening of Theiler's murine encephalomyelitis virus antibodies in mice colonies

Juan M. Laborde, Cecilia Carbone, Santiago G. Corva and Cecilia M. Galosi1

Correspondence: 1Corresponding Author: Cecilia M. Galosi, Department of Virology, Faculty of Veterinary Sciences, National University of La Plata, 60 and 118, PO Box 296, 1900 La Plata, Buenos Aires, Argentina. cmgalosi{at}fcv.unlp.edu.ar, cmgalosi{at}yahoo.com

The current study demonstrates the ability of an indirect enzyme-linked immunosorbent assay (iELISA) to detect antibodies against Theiler's murine encephalomyelitis virus in mice colonies. The antigen was produced from infected baby hamster kidney (BHK)-21 cells and treated with 1% Nonidet P40 in saline buffer. Control antigen was prepared following the same procedure using uninfected BHK-21 cells. The optimal antigen and serum dilutions were established. The reaction was revealed using an anti-mouse–horseradish peroxidase conjugate and 2,2'-Azino-bis(3-ethylbenzthiazoline-6-sulfonic acid). Optimized iELISA was validated by detection of antibodies in known positive and negative serum samples before testing the samples of unknown status. Performance of the iELISA was compared with the indirect fluorescent antibody test, and the cutoff value was determined by receiver operating curve. Indirect ELISA showed 100% sensitivity, 99.38% specificity, and 97.78% predictive positive value. The antigen used is easy to produce, and no special equipment is required. The iELISA developed is simple and provides a rapid and less costly tool for diagnosis and research.

Key Words: Diagnosis • enzyme-linked immunosorbent assay • Theiler's murine encephalomyelitis virus






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