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Detection of PMV-1 specific antibodies with a monoclonal antibody blocking enzyme-linked immunosorbent assay
G Czifra1, M Nilsson, D J Alexander
1Division of Poultry, The National Veterinary Institute, Uppsala, Sweden.
Abstract:
A highly reproducible monoclonal antibody (Mab) blocking ELISA (B-ELISA) has been developed and evaluated for the detection of NDV-specific antibodies. The Mab utilised is specific for a conserved PMV-1 serotype-specific epitope, as demonstrated by the indirect immunoperoxidase test. It reacted with all strains representing different serogroups within the PMV-1 serotype, but not with any strain belonging to other PMV serotypes. Sensitivity and specificity of the B-ELISA were compared with the haemagglutina-tion inhibition test (HI). Blocking and HI antibodies were detected in sera of chickens 8 days post-experimental infection. The B-ELISA proved consistently more sensitive than the HI test. In another survey, 62 sera from experimentally vaccinated chickens were tested; 95.2% proved positive by B-ELISA, 85.5% by indirect ELISA and 74% by HI test. When 504 field sera from vaccinated chickens and turkeys were tested, 98% were positive by B-ELISA, and 69% by HI. The specificity was evaluated by testing 1066 samples from NDV-free flocks, all of which proved negative by both methods. Other advantages of the B-ELISA include easy standardization and quality control, and ability to test sera from any species (including exotic or wild birds as well as mammals). The use of low dilution serum or egg-yolk samples makes the test quick and easy to perform and suitable for large-scale screening.
Insights
A new monoclonal antibody blocking ELISA (B-ELISA) offers highly sensitive and specific detection of Newcastle disease virus (NDV) antibodies. This B-ELISA is superior to traditional methods for large-scale screening in various avian and mammalian species.
Area of Science:
- Veterinary Immunology
- Avian Pathology
- Diagnostic Assays
Background:
- Newcastle disease virus (NDV) poses a significant threat to poultry health and global trade.
- Accurate and sensitive detection of NDV-specific antibodies is crucial for disease surveillance and vaccine efficacy assessment.
- Existing diagnostic methods like the haemagglutination inhibition (HI) test have limitations in sensitivity and scope.
Purpose of the Study:
- To develop and evaluate a highly reproducible monoclonal antibody blocking ELISA (B-ELISA) for detecting NDV-specific antibodies.
- To compare the sensitivity and specificity of the B-ELISA against the traditional HI test.
- To assess the applicability of the B-ELISA for large-scale screening across different species.
Main Methods:
- Development of a monoclonal antibody (Mab) specific for a conserved PMV-1 serotype-specific epitope.
- Evaluation of the Mab using indirect immunoperoxidase test against various PMV serotypes.
- Comparison of B-ELISA performance with HI test using experimentally infected and vaccinated chicken sera, and field sera from chickens and turkeys.
- Specificity assessment using sera from NDV-free flocks.
Main Results:
- The Mab demonstrated specificity for PMV-1 serotype strains, reacting with all tested PMV-1 strains but not other PMV serotypes.
- The B-ELISA consistently showed higher sensitivity than the HI test in detecting antibodies post-infection and in vaccinated populations.
- In surveys, B-ELISA detected antibodies in 95.2% of experimentally vaccinated chickens and 98% of field sera from vaccinated chickens and turkeys, compared to lower percentages for HI.
- 1066 samples from NDV-free flocks were negative by both B-ELISA and HI, confirming high specificity.
Conclusions:
- The developed B-ELISA is a highly sensitive and specific method for detecting NDV-specific antibodies.
- The B-ELISA offers advantages over the HI test, including superior sensitivity, ease of standardization, and suitability for large-scale screening.
- The B-ELISA is adaptable for testing sera from diverse species, including wild birds and mammals, enhancing its utility in disease monitoring.
