title: Two quantitative real-time PCR assays for the detection of penaeid shrimp and blue crab, crustacean shellfish allergens.
abstract: Food allergen detection methods must be able to specifically detect minute quantities of an allergenic food in a complex food matrix. One technique that can be used is real-time PCR. For the work described here, real-time PCR assays were developed to detect penaeid shrimp and blue crab, crustacean shellfish allergens. The method was tested using shrimp meat and crab meat spiked into several types of foods, including canned soups, deli foods, meat, seafood, and prepared seafood products. Foods were spiked with either shrimp or crab at levels ranging from 0.1 to 10⁶ parts per million (ppm) and analyzed either raw or cooked by a variety of methods. Real-time PCR data were used to generate linear standard curves, and assays were evaluated with respect to linear range and reaction efficiency. Results indicate that both assays performed well in a variety of food types. High reaction efficiencies were achieved across a linear range of 6-8 orders of magnitude. Limits of detection were generally between 0.1 and 1 ppm. Cooking methods used to simulate thermal processing of foods had little effect on assay performance. This work demonstrates that real-time PCR can be a valuable tool in the detection of crustacean shellfish.
authors.affiliation: Division of Bioanalytical Chemistry, Office of Regulatory Science, Center for Food Safety and Applied Nutrition, U.S. Food and Drug Administration , 5100 Paint Branch Parkway, Mailstop HFS-716, College Park, Maryland 20740, United States.
authors.forename: Anne C
authors.initials: AC
authors.lastname: Eischeid
authors.forename: Bang-hyun
authors.initials: BH
authors.lastname: Kim
authors.forename: Sasha M
authors.initials: SM
authors.lastname: Kasko
mesh_terms: D000485:Allergens / Q000009:adverse effects / Q000032:analysis* / Q000737:chemistry; D000818:Animals; D060829:Arthropod Proteins / Q000009:adverse effects / Q000032:analysis* / Q000737:chemistry; D003386:Brachyura / Q000737:chemistry*; D003296:Cooking; D004044:Dietary Proteins / Q000009:adverse effects / Q000032:analysis / Q000737:chemistry; D057140:Fast Foods / Q000009:adverse effects / Q000032:analysis*; D005512:Food Hypersensitivity / Q000209:etiology / Q000517:prevention & control; D005513:Food Inspection / Q000379:methods*; D006801:Humans; D060885:Multiplex Polymerase Chain Reaction; D033561:Penaeidae / Q000737:chemistry*; D055550:Protein Stability; D060888:Real-Time Polymerase Chain Reaction; D012758:Shellfish / Q000009:adverse effects / Q000032:analysis*
chemical_list: D000485:Allergens; D060829:Arthropod Proteins; D004044:Dietary Proteins