Simplex Immunodotblot for detecting dog meat contaminants in foodstuffs

This study is an initial effort to develop a prototype kit based on the simplex immunodot blot method to detect dog meat contamination in food products. The simplex immunodotblot is an immunoassay technique that identifies specific proteins or antigens through antibody-antigen interactions, targeting a single contaminant in one reaction. The antigen utilized was dog meat extract (DME) prepared in the laboratory, while the primary antibodies (Ab) against DME were produced in-house using laying hens. Test samples were applied in varying concentrations (5, 10, and 20 µg per spot) onto comb-shaped polyester plastic sheets. A commercial secondary antibody against IgY, labeled with horseradish peroxidase (HRP), served as the detector. The reaction was visually inspected; in cases where dog meat was present, a blue or black spot appeared due to the reaction between the antigen, antibody, enzyme, and substrate. The study demonstrated the successful development of the simplex immunodotblot, which was able to detect dog meat visually at concentrations as low as 5 µg in a laboratory setting. However, further research is required, particularly to adapt the prototype kit for halal food authentication purposes.