Lab-on-a-chip (LOC) devices have been developed for nucleic acid analysis by integrating complex laboratory functions onto a miniaturized chip, enabling rapid, cost-effective, and highly sensitive on-site testing. This review examines the application of LOC technology in food safety, specifically in the context of nucleic acid-based analyses for detecting pathogens and contaminants. We focus on microfluidic-based LOC devices that optimize nucleic acid extraction and purification on the chip or amplification and detection processes based on isothermal amplification and polymerase chain reaction. We also explore advancements in integrated LOC devices that combine nucleic acid extraction, amplification, and detection processes within a single chip to minimize sample preparation time and enhance testing accuracy. The review concludes with insights into future trends, particularly the development of portable LOC technologies for rapid and efficient nucleic acid testing in food safety.