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Journal of Clinical Microbiology, March 2003, p. 943-947, Vol. 41, No. 3
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.3.943-947.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Competitive Enzyme-Linked Immunosorbent Assay Based on Monoclonal Antibody and Recombinant Hemagglutinin for Serosurveillance of Rinderpest Virus

G. J. Renukaradhya,1 K. B. Suresh,1 M. Rajasekhar,1 and M. S. Shaila2*

Project Directorate on Animal Disease Monitoring and Surveillance, Hebbal, Bangalore-560 024,1 Department of Microbiology and Cell Biology, Indian Institute of Science, Bangalore-560 012, India2

Received 29 April 2002/ Returned for modification 16 August 2002/ Accepted 15 December 2002

A competitive enzyme-linked immunosorbent assay (C-ELISA) which detects antibodies unique to rinderpest virus (RPV) has been developed. This test can differentiate antibodies against RPV and those against peste des petits ruminants virus. The recombinant RPV hemagglutinin (H)-protein C-ELISA (recH C-ELISA) is based on the ability of a well-characterized monoclonal antibody (MAb) produced with the soluble, secreted form of the H protein (Sec H protein) of RPV made in a baculovirus expression system to compete with the binding of RPV antibodies in the serum of vaccinated or infected, recovered animals to the Sec H protein. The B-cell epitope recognized by the MAb corresponds to amino acids 575 to 583 on the H protein, which is not present on the antigenically closely related peste des petits ruminants virus hemagglutinin-neuraminidase protein. Initially, a positive-negative threshold cutoff value for percent inhibition of 34 was established with 500 known RPV-negative serum samples. The recH C-ELISA was developed with the enzyme immunoassay software of a commercial RPV C-ELISA kit. Comparative analysis of the test results for 700 serum samples obtained with the commercial kit gave a sensitivity of 112.4% and a specificity of 72.4%. Variations in percent inhibition values were observed for the two assay systems. These variations may have been due to the undefined amount of antigen present in the commercial kit as well as the use of a different MAb. The recH C-ELISA detected more positive serum samples compared to the number detected by the commercial kit, with the results confirmed by a virus neutralization test. Thus, recH C-ELISA is a sensitive tool for RPV serosurveillance in disease eradication programs.

* Corresponding author. Mailing address: Department of Microbiology and Cell Biology, Indian Institute of Science, Bangalore-560 012, India. Phone: 91-80-394 2702. Fax: 91-80-360 2697. E-mail: shaila{at}mcbl.iisc.ernet.in.

Journal of Clinical Microbiology, March 2003, p. 943-947, Vol. 41, No. 3
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.3.943-947.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Balamurugan, V., Sen, A., Saravanan, P., Rasool, T. J., Yadav, M. P., Bandyopadhyay, S. K., Singh, R. K. (2006). Development and Characterization of a Stable Vero Cell Line Constitutively Expressing Peste des Petits Ruminants Virus (PPRV) Hemagglutinin Protein and Its Potential Use as Antigen in Enzyme-Linked Immunosorbent Assay for Serosurveillance of PPRV. CVI 13: 1367-1372 [Abstract] [Full Text]  


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