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Detection of peste des petits ruminants virus antigen using immunofiltration and antigen-competition ELISA methods

Raj, Dhinakar G and Rajanathan, TMC and Kumar, Senthil C and Ramathilagam, G and Hiremath, Geetha and Shaila, MS (2008) Detection of peste des petits ruminants virus antigen using immunofiltration and antigen-competition ELISA methods. In: Veterinary Microbiology, 129 (3-4). pp. 246-251.

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Abstract

Peste des petits ruminants (PPR) is one of the most economically important diseases affecting sheep and goats in India. An immunofiltration-based test has been developed using either mono-specific serum/monoclonal antibodies (mAb) prepared against a recombinant truncated nucleocapsid protein of rinderpest virus (RPV) cross-reactive with PPR virus. This method consists of coating ocular swab eluate from suspected animals onto a nitrocellulose membrane housed in a plastic module, which is allowed to react with suitable dilutions of a mAb or a mono-specific polyclonal antibody. The antigen–antibody complex formed on the membrane is then detected by protein A-colloidal gold conjugate, which forms a pink colour. In the immunofiltration test, concordant results were obtained using either PPRV mAb or mono-specific serum. Another test, an antigen-competition ELISA which relies on the competition between plate-coated recombinant truncated ‘N’ protein of RPVand the PPRV ‘N’ protein present in ocular swab eluates (sample) for binding to the mono-specific antibody against N protein of RPV (in liquid phase) was developed. The cut-off value for this test was established using reverse transcription polymerase chain reaction (RT-PCR) positive and negative oculo-nasal swab samples. Linear correlation between percent inhibition (PI) values in antigen-competition ELISA and virus infectivity titres was 0.992. Comparison of the immunofiltration test with the antigen-competition ELISA yielded a sensitivity of 80% and specificity of 100%. These two tests can serve as a screening (immunofiltration) and confirmatory (antigen-competition ELISA) test,respectively, in the diagnosis of PPR in sheep or goats.

Item Type: Journal Article
Additional Information: Copyright of this article belongs to Elsevier.
Keywords: Peste des petits ruminants;Immunofiltration;Antigen-competition ELISA;RT-PCR;Diagnosis.
Department/Centre: Division of Biological Sciences > Microbiology & Cell Biology
Date Deposited: 10 Jul 2008
Last Modified: 19 Sep 2010 04:47
URI: http://eprints.iisc.ernet.in/id/eprint/14915

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