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Protein and DNA-based assays as complementary tools for fish allergen detection*

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 authentication of eight commonly consumed fishes in food and...

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Autores principales: Kuehn, A., Hilger, C., Graf, T., Hentges, F.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Dustri-Verlag Dr. Karl Feistle 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6040006/
https://www.ncbi.nlm.nih.gov/pubmed/30402610
http://dx.doi.org/10.5414/ALX01485E
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author Kuehn, A.
Hilger, C.
Graf, T.
Hentges, F.
author_facet Kuehn, A.
Hilger, C.
Graf, T.
Hentges, F.
author_sort Kuehn, A.
collection PubMed
description 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 authentication 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.
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spelling pubmed-60400062018-11-06 Protein and DNA-based assays as complementary tools for fish allergen detection* Kuehn, A. Hilger, C. Graf, T. Hentges, F. Allergol Select Research Article 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 authentication 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. Dustri-Verlag Dr. Karl Feistle 2017-08-04 /pmc/articles/PMC6040006/ /pubmed/30402610 http://dx.doi.org/10.5414/ALX01485E Text en © Dustri-Verlag Dr. K. Feistle http://creativecommons.org/licenses/by/2.5/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Kuehn, A.
Hilger, C.
Graf, T.
Hentges, F.
Protein and DNA-based assays as complementary tools for fish allergen detection*
title Protein and DNA-based assays as complementary tools for fish allergen detection*
title_full Protein and DNA-based assays as complementary tools for fish allergen detection*
title_fullStr Protein and DNA-based assays as complementary tools for fish allergen detection*
title_full_unstemmed Protein and DNA-based assays as complementary tools for fish allergen detection*
title_short Protein and DNA-based assays as complementary tools for fish allergen detection*
title_sort protein and dna-based assays as complementary tools for fish allergen detection*
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6040006/
https://www.ncbi.nlm.nih.gov/pubmed/30402610
http://dx.doi.org/10.5414/ALX01485E
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