Staphylococcal enterotoxin is a very common cause of food poisoning. Conventional detection methods for the toxin including enzyme–linked immunosorbent assays (ELISAs), chemiluminescence (ECL), and polymerase chain reaction (PCR) assays require a lot of time, efforts, and expert technicians. Lateral flow strip kits have shown great potential for the rapid detection of foodborne pathogens. The lateral flow strip kit is widely used in clinical settings because it is easy to use, fast, and cost effective. A typical lateral flow strip kit uses colloidal gold to generate a visual signal. However, the lateral flow strip kit based on colloidal gold has limited sensitivity to fulfill food safety regulation requirements. This study was performed to develop a rapid test kit for pathogenic staphylococcal enterotoxin B (SEB) in food samples. The rapid detection kit was fabricated based on a nitrocellulose lateral-flow strip. Cellulose nano beads and SEB antibodies were used as the tag and receptor, respectively, to improve the detection performance. Manually spotted SEB antibody and anti-rabbit antibody on the surface of the nitrocellulose membrane were used as test and control spots, respectively. The feasibility of the rapid test kit to detect SEB in samples was evaluated. The sensitivity of the kit was 10 ng/mL SEB spiked in PBS. Additionally, the rapid test kit could detect 1 ng/mL of SEB in chicken meat extract.