The invention discloses a preparation method and an application method of a nucleic acid rapid detection kit for detection of rabbit source (Leporidae) components in food and feeds, and belongs to the field of molecular biology and immunology. According to the preparation method and the application method, a high sensitivity and high specificity method of polymerase chain reaction for nucleic acid detection and an immune colloidal gold rapid detection technology of immunological detection can be combined, a unique primer is designed, the primer is marked, extracted target DNA is specifically amplified, and an amplified product is combined with a gold labeled antibody fixed on a test paper strip in a developing solution for formation of a stable and visible detection band and a quality control band, so that fast and accurate detection of the rabbit source components in the food and feeds can be realized.