The compound 1,1,1-trichloro-2-(p-chlorophenyl)-2-(o-chlorophenyl) ethane (o,p’-DDT) has historically been used as a pesticide, but is now recognized as an endocrine-disrupting chemical with the potential to accumulate in the food chain and cause adverse effects on wildlife and humans. Given its persistence in the environment, the detection of o,p'-DDT residues in food products has become a matter of significant concern for food safety and public health. Aptamer are short, single-stranded nucleic acid (DNA or RNA) molecules that have the ability to bind to a specific target molecule with high affinity and specificity. In this study, employing the powerful SELEX (Systematic Evolution of Ligands by EXponential Enrichment) technique, we have successfully identified aptamers that specifically bind to o,p'-DDT. From an initial pool of seventeen candidates, we narrowed down this number to five for preliminary assessment using the SYBR Green I assay. One of the candidates, DDT_13, exhibited a pronounced fluorescent response to o,p'-DDT. Subsequent analysis revealed DDT_13's dissociation constant (Kd) to be 412.3 ± 124.6 nM, indicating a robust binding affinity. Furthermore, DDT_13 displayed negligible non-specific interactions with other small molecules, emphasizing its exceptional specificity. The aptamer's efficacy in analyzing food samples underscores its promising bioactivity, positioning it as a novel tool for o,p'-DDT detection in the context of food safety monitoring and risk evaluation initiatives.