Detection of crustacean DNA and species identification using a PCR-restriction fragment length polymorphism method

The detection of potentially allergenic proteins, such as those derived from crustaceans, in food products is a major concern for the food processing industry. A PCR-restriction fragment length polymorphism (PCR-RFLP) method was designed to detect the presence of crustacean DNA in food products and to determine the species source of the DNA. This PCR assay amplifies an approximately 205-bp fragment of the 16S rRNA gene in crustacean species, including shrimp, crab, lobster, and crawfish. This reaction will not amplify DNA derived from mammals, such as cow and sheep. After amplification, the PCR product is digested with differential restriction endonucleases to determine the species source of the crustacean DNA. The specificity of this assay was demonstrated using four species of shrimp, three species of crab, and two species of lobster and crawfish. This assay is sensitive enough to detect crustacean DNA in a raw meat mixture containing <0.1% shrimp.