Peer-reviewed veterinary case report
Development of an indirect ELISA with artificially synthesized N protein of PPR virus.
- Journal:
- Intervirology
- Year:
- 2012
- Authors:
- Zhang, Guo-Rui et al.
- Affiliation:
- College of Veterinary Medicine · China
Plain-English summary
Researchers developed a new test to detect antibodies against the peste des petits ruminants virus (PPRV), which affects sheep and goats. They created a specific protein from the virus in the lab and used it in a test that can identify infected animals by checking their blood samples. This new test was found to be very accurate, correctly identifying 96.7% of positive cases and 96.1% of negative cases when compared to an existing test. Overall, the new test appears to work very well for detecting PPRV in small ruminants.
Abstract
The full-length gene encoding the nucleocapsid (N) protein of the virus (PPRV) responsible for an outbreak of peste des petits ruminants in Tibet in 2007 was synthesized in two stages using overlapping PCR without the need for viral genomic cDNA as template. The full-length N gene was successfully expressed in Escherichia coli, and the purified gene product bound to monoclonal antibody raised against PPRV N protein. Furthermore, it was able to replace recombinant B-N antigen as the coating antigen in a commercial ELISA kit prepared with another PPRV strain. Recombinant protein was employed as the coating antigen to develop an indirect ELISA for PPRV antibody detection in the sera of infected small ruminants. Antibody detection was optimal at a 1:200 serum dilution and an antigen concentration of 3.2 μg/ml, and the positive threshold (cutoff) value of the assay was 2.18. Analysis of 697 serum samples revealed the sensitivity and specificity of the indirect ELISA to be 96.7 and 96.1%, respectively, compared with a commercially available ELISA test.
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Search related cases →Original publication: https://pubmed.ncbi.nlm.nih.gov/21242661/