Accurate data on mycotoxin contamination and exposure patterns is crucial in contributing to food safety policies and public health interventions in low-resource nations in the developing world. In such settings, with inadequate research infrastructures and high-performance analytical tools, the enzyme-linked immunosorbent assay (ELISA) technique remains the most widely used technique to measure regulated mycotoxins (e.g., aflatoxins and fumonisins) in food, feed, blood, and urine samples. The results of ELISA technique alone or/in combination with food frequency intake survey data may generally be employed to estimate dietary exposure levels of populations and individuals. This report provides collection of ELISA-based mycotoxin detection protocols (including strengths and weaknesses) with guides for reasonable results.