Food allergies are a potential food safety and public health concern worldwide. To assure the safety of people who experience allergic reactions, it is necessary to establish effective and reliable methods for rapid detection of food allergens. This paper reports an innovative method for the rapid detection and analysis of porcine serum albumin (PSA), known as a major allergen in pork, based on a surface plasmon resonance (SPR) biosensor. The antibodies known to have a high bioactivity against PSA were verified by competitive indirect-ELISA and then immobilized on the SPR sensor surface, thus allowing them to capture PSA. The developed SPR demonstrated a linear range from 1.0 to 450 ng/mL for the measurement of PSA with a detection limit of 19.81 ng/mL. Within-day RSD (1.97-4.02%) and between-day RSD (1.88-4.15%) were no more than 5%. The SPR was evaluated for analysis of six commercial food samples and showed almost perfect agreement between the results obtained by ELISA test kits without significant differences (P > 0.05). Therefore, this assay permits accurate, specific, and sensitive detection of PSA in pork and pork products.