Protein- und DNS-basierte methoden als komplementäre methoden zum nachweis von fischallergenen in nahrungsmitteln

Translated title of the contribution: Protein- and DNA-based assays as complementary tools for fish allergen detection

Annette Kuehn*, C. Hilger, T. Graf, F. Hentges

*Corresponding author for this work

Research output: Contribution to journalArticleResearchpeer-review

9 Citations (Scopus)

Abstract

Background: Fish is one of the most important, allergenic foods worldwide. Parvalbumin is the well characterized, major allergen in fish muscle. In this study, we developed a protein- and a DNA-based method for the sensitive detection and authentification of eight commonly consumed fishes in food and compared their applicability. Methods: Fish parvalbumins were purified. Polyclonal, anti-parvalbumin antibodies were raised in rabbits and mice. Protein extracts from food were analyzed by quantitative ELISA. Parvalbumin genes were cloned and sequenced for the design of parvalbumin gene-specific PCR-primers. DNA extracted from food was subjected to specific PCR. Results: Increasing parvalbumin contents were quantified by ELISA in fresh fish, in the order of tuna < mackerel < cod < salmon/trout < redfish < carp < herring. The parvalbumin content of processed fish was up to 67% lower than in fresh fish. In spiked food samples, 1 to 15 ppm fresh fish and 30 to 170 ppm processed fish were still detectable by ELISA. The eight fishes were identified by specific PCR using 0.2 to 10 ng fish DNA. PCRs detected still 3 ppm fresh fish and 30 to 150 ppm processed fish in spiked samples. Conclusions: Both the protein- and the DNA-based method have sufficient sensitivity to protect fish-allergic consumers. The ELISA allows allergen quantification, while the PCR identifies the fish present in the food. The detection limits of both methods vary depending on different factors. Both methods need to be carefully validated for each fish and fish product when used in detection assays.

Translated title of the contributionProtein- and DNA-based assays as complementary tools for fish allergen detection
Original languageGerman
Pages (from-to)343-350
Number of pages8
JournalAllergologie
Volume35
Issue number7
DOIs
Publication statusPublished - Jul 2012

Keywords

  • Allergen detection
  • DNA
  • ELISA
  • Fish allergy
  • PCR
  • Parvalbumin
  • Protein

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