This chapter provides a comprehensive overview of the Direct Antigen Capture Enzyme-Linked Immunosorbent Assay (DAC ELISA), a sensitive and specific technique for detecting and quantifying antigens in various biological samples. The chapter outlines the step-by-step protocol for conducting DAC ELISA, including sample preparation, coating of plates with capture antibodies, and the detection process using enzyme-labeled antibodies. Key factors such as antibody specificity, incubation times, and washing procedure influence the assay. The Direct Antigen Capture Enzyme-Linked Immunosorbent Assay (DAC ELISA) is vital for its high sensitivity and specificity in detecting target antigens, making it a cornerstone in clinical diagnostics and research. Its ability to deliver rapid results is crucial in time-sensitive situations, such as diagnosing infectious diseases. Versatile in application, DAC ELISA is utilized across various fields. The assay provides quantitative data, essential for tracking disease progression and therapeutic responses, while remaining cost-effective and straightforward to implement, making it accessible to a wide range of laboratories. Its adaptability allows for customization based on specific antigens, enhancing its utility in diverse research scenarios. Ultimately, DAC ELISA contributes significantly to pathogen detection and diagnostics by enabling the early detection of pathogens and toxins, thereby playing a critical role in monitoring and preventing diseases. This chapter serves as a practical guide for researchers and laboratory technicians seeking to implement DAC ELISA in their work.