Development and Implementation of a Sandwich ELISA Prototype for the Detection of Brucella Antigens in Cheese Samples

Abstract

Brucellosis remains one of the most important zoonotic diseases worldwide, posing a serious threat to both public health and animal production. Unpasteurized dairy products represent a major route of transmission. Furthermore, even in regions where the disease has been eradicated, individuals may remain at risk due to travel or the illegal importation of contaminated products. Effective control therefore relies on rapid, accurate, and practical diagnostic methods. In this study, a sandwich enzyme-linked immunosorbent assay (ELISA) prototype was developed to detect Brucella antigens in cheese irrespective of bacterial viability. The assay was evaluated using 150 fresh cheese samples collected from the Central and Siverek districts of S & cedil;anlıurfa province, Türkiye. The developed test demonstrated 100% specificity and 66% sensitivity. Overall, 17.3% of the samples were positive for Brucella antigens, with similar positivity rates observed across both districts. These findings indicate a considerable public health risk associated with contaminated fresh cheese. The newly developed sandwich ELISA may serve as a practical tool for antigen detection and support brucellosis surveillance and control efforts.