Quantitative sandwich ELISA for the determination of fish in foods.
Allergy to fish represents one of the most prevalent causes for severe food-allergic reactions. Therefore, food authorities in different countries have implemented mandatory labeling of fish in pre-packed foods. Detection of fish proteins in food has previously been based on the use of patient serum. In the present study, a novel sandwich enzyme-linked immunosorbent assay (ELISA) for the quantitation of fish in food matrixes has been developed and validated, using a polyclonal rabbit anti-cod parvalbumin antibody for capture and a biotinylated conjugate of the same antibody for detection. By employing the ubiquitous muscle protein parvalbumin as target the method succeeds to detect a variety of fish. However, the ELISA is specific for fish and does not cross-react with other species. Recoveries ranged from 68-138% in typical food matrixes, while the intra- and inter-assay precisions were <12% and <19%, respectively. The sensitivity of the cod parvalbumin ELISA with a limit of detection of 0.01 mg parvalbumin/kg food, about 5 mg fish/kg food, seems sufficient to detect fish protein traces in foods at levels low enough to minimize the risk for fish allergic consumers.
Faeste
Christiane K
CK
National Veterinary Institute, Post Office Box 8156 Dep., N-0033 Oslo, Norway. christiane.faste@vetinst.no
Plassen
Christin
C
D000485:Allergens / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification; D000818:Animals; D000906:Antibodies; D000918:Antibody Specificity; D019921:Biotinylation; D002138:Calibration; D004797:Enzyme-Linked Immunosorbent Assay / Q000592:standards; D029941:Fish Proteins / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification; D005504:Food Analysis / Q000379:methods* / Q000592:standards; D005506:Food Contamination; D005512:Food Hypersensitivity / Q000517:prevention & control; D048389:Gadus morhua; D006801:Humans; D010320:Parvalbumins / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification; D015203:Reproducibility of Results; D013045:Species Specificity
D000485:Allergens; D000906:Antibodies; D029941:Fish Proteins; D010320:Parvalbumins