Development and evaluation of egg yolk-derived antibodies for ELISA diagnostics of African Animal Trypanosomiasis

Abstract

African animal trypanosomiasis (AAT) represents a significant challenge to livestock development in Africa. Reliable and practical techniques are required for the disease's early detection and management. One of the most commonly used tests for this purpose is the Enzyme-Linked Immunosorbent Assay (ELISA). This study sought to create a protocol for producing labelled antibodies from egg yolk. IgG was purified from serum from cattle that were hyperimmune to Trypanosoma brucei brucei before being used to immunise chickens. IgY antibodies were extracted from eggs, labelled with peroxidase, and tested for activity against commercial products. The results revealed that IgY levels were consistently higher than IgG levels, and the experimental antiserum had high diagnostic potential. We also calculated the ratios of commercial and individual egg yolk antisera. The findings allowed us to rank the diagnostic potential of the experimental antisera, with detection rates of 47.33 % for positive samples and 41.47 % for negative samples. Our results show that the experimental antiserum detects target antibodies with comparable accuracy and statistical significance (p < 0.001). Furthermore, the production method based on laying chickens proved to be simple, effective, and economical. This locally synthesised antiserum provides a viable alternative to expensive commercial options, paving the way for more widespread use in serodiagnosis. Further refinement and validation of this methodology could result in the development of a standardised protocol for large-scale production, offering a cost-effective and ethically sound alternative to antiserum production and facilitating wider adoption of ELISA diagnostics in resource-constrained settings.