Shrimp is the popular food within high nutritional values. Specially, the shrimp is consumed as a raw material in various processed foods worldwide. However, shrimp can commonly trigger the seafood allergic reactions that threaten the lives of people. In this study, the PCR assay was developed to detect the presence of tropomyosin - a shrimp-derived major allergen. For allergen detection, a speciﬁc primer pair, Tropo-F and Tropo-R, was designed based on the shrimp tropomyosin gene. The specificity of primer was accessed by in silico study and initially applied in a sample of Litopenaeus vannamei and commercial shrimp-derived processed food. As the results, an 81-bp band was observed in the sample of Litopenaeus vannamei and other commercial shrimp-derived component foods, and no band was observed in non-shrimp samples. The specificity was confirmed by sequencing, which indicated that the PCR products were homologous to Litopenaeus vannamei and Penaeus monodon. In summary, these results indicated that PCR assay with Tropo-F and Tropo-R primers could successfully identify the tropomyosin in processed food, and could be applied to a large number of samples in the reality. To increase the value of this PCR assay, the sensitivity of this assay is necessary to be evaluated in our near studies.