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Competitive enzyme-linked immunosorbent assay based on monoclonal antibody and recombinant hemagglutinin for serosurveillance of rinderpest virus
- Source :
- IndraStra Global.
- Publication Year :
- 2003
- Publisher :
- American Society for Microbiology, 2003.
-
Abstract
- 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.
- Subjects :
- Microbiology (medical)
Paramyxoviridae
medicine.drug_class
Hemagglutinins, Viral
Enzyme-Linked Immunosorbent Assay
Spodoptera
Monoclonal antibody
Rinderpest virus
Epitope
Viral Proteins
Virology
medicine
Animals
Mononegavirales
Cells, Cultured
Glycoproteins
Microbiology & Cell Biology
biology
medicine.diagnostic_test
Antibodies, Monoclonal
Hemagglutinin
biology.organism_classification
Molecular biology
Recombinant Proteins
Hemagglutinins
Immunoassay
biology.protein
Cattle
Reagent Kits, Diagnostic
Antibody
Subjects
Details
- Language :
- English
- ISSN :
- 23813652
- Database :
- OpenAIRE
- Journal :
- IndraStra Global
- Accession number :
- edsair.doi.dedup.....b07b2454f9cf7198f50338029780222c