A method to detect allergenic fish, specifically cod and pollock, using quantitative real‐time PCR and COI DNA barcoding sequences
Abstract BACKGROUND Fish are one of eight major allergens defined in the US Food Allergen Labeling and Consumer Protection Act, and cod and pollock are two of the major fish allergens. This paper describes development and validation of a method to detect cod and pollock in complex food matrices usin...
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crwiley:10.1002/jsfa.9466 2024-09-15T17:55:33+00:00 A method to detect allergenic fish, specifically cod and pollock, using quantitative real‐time PCR and COI DNA barcoding sequences Eischeid, Anne C 2018 http://dx.doi.org/10.1002/jsfa.9466 https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1002%2Fjsfa.9466 https://onlinelibrary.wiley.com/doi/pdf/10.1002/jsfa.9466 en eng Wiley http://onlinelibrary.wiley.com/termsAndConditions#vor Journal of the Science of Food and Agriculture volume 99, issue 5, page 2641-2645 ISSN 0022-5142 1097-0010 journal-article 2018 crwiley https://doi.org/10.1002/jsfa.9466 2024-08-06T04:17:23Z Abstract BACKGROUND Fish are one of eight major allergens defined in the US Food Allergen Labeling and Consumer Protection Act, and cod and pollock are two of the major fish allergens. This paper describes development and validation of a method to detect cod and pollock in complex food matrices using real‐time polymerase chain reaction (PCR). Mitochondrial cytochrome oxidase I (COI) sequences obtained through DNA barcoding were used to design a single set of primers and probe which detected three species in the genus Gadus : Atlantic cod, Pacific cod, and walleye pollock. RESULTS Cod spiked into three different food matrices (cooking oil, clam chowder, and hushpuppy mix) yielded high linearity, dynamic range spanning six orders of magnitude, and lower limits of detection at 1–10 ppm (ppm; mg kg −1 ). Frying had an adverse effect on the lower limit of detection, but not on linearity. CONCLUSIONS This work shows that COI DNA barcoding sequences can be used to effectively design real‐time PCR assays for detection of food allergens in complex matrices. While full‐length DNA barcodes distinguish individual species, the PCR assay designed here detected three different species. This is likely because real‐time PCR assays are tolerant to basepair mismatches and do not utilize the full length of the DNA barcode. Published 2018. This article is a U.S. Government work and is in the public domain in the USA. Article in Journal/Newspaper atlantic cod Wiley Online Library Journal of the Science of Food and Agriculture 99 5 2641 2645 |
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Abstract BACKGROUND Fish are one of eight major allergens defined in the US Food Allergen Labeling and Consumer Protection Act, and cod and pollock are two of the major fish allergens. This paper describes development and validation of a method to detect cod and pollock in complex food matrices using real‐time polymerase chain reaction (PCR). Mitochondrial cytochrome oxidase I (COI) sequences obtained through DNA barcoding were used to design a single set of primers and probe which detected three species in the genus Gadus : Atlantic cod, Pacific cod, and walleye pollock. RESULTS Cod spiked into three different food matrices (cooking oil, clam chowder, and hushpuppy mix) yielded high linearity, dynamic range spanning six orders of magnitude, and lower limits of detection at 1–10 ppm (ppm; mg kg −1 ). Frying had an adverse effect on the lower limit of detection, but not on linearity. CONCLUSIONS This work shows that COI DNA barcoding sequences can be used to effectively design real‐time PCR assays for detection of food allergens in complex matrices. While full‐length DNA barcodes distinguish individual species, the PCR assay designed here detected three different species. This is likely because real‐time PCR assays are tolerant to basepair mismatches and do not utilize the full length of the DNA barcode. Published 2018. This article is a U.S. Government work and is in the public domain in the USA. |
format |
Article in Journal/Newspaper |
author |
Eischeid, Anne C |
spellingShingle |
Eischeid, Anne C A method to detect allergenic fish, specifically cod and pollock, using quantitative real‐time PCR and COI DNA barcoding sequences |
author_facet |
Eischeid, Anne C |
author_sort |
Eischeid, Anne C |
title |
A method to detect allergenic fish, specifically cod and pollock, using quantitative real‐time PCR and COI DNA barcoding sequences |
title_short |
A method to detect allergenic fish, specifically cod and pollock, using quantitative real‐time PCR and COI DNA barcoding sequences |
title_full |
A method to detect allergenic fish, specifically cod and pollock, using quantitative real‐time PCR and COI DNA barcoding sequences |
title_fullStr |
A method to detect allergenic fish, specifically cod and pollock, using quantitative real‐time PCR and COI DNA barcoding sequences |
title_full_unstemmed |
A method to detect allergenic fish, specifically cod and pollock, using quantitative real‐time PCR and COI DNA barcoding sequences |
title_sort |
method to detect allergenic fish, specifically cod and pollock, using quantitative real‐time pcr and coi dna barcoding sequences |
publisher |
Wiley |
publishDate |
2018 |
url |
http://dx.doi.org/10.1002/jsfa.9466 https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1002%2Fjsfa.9466 https://onlinelibrary.wiley.com/doi/pdf/10.1002/jsfa.9466 |
genre |
atlantic cod |
genre_facet |
atlantic cod |
op_source |
Journal of the Science of Food and Agriculture volume 99, issue 5, page 2641-2645 ISSN 0022-5142 1097-0010 |
op_rights |
http://onlinelibrary.wiley.com/termsAndConditions#vor |
op_doi |
https://doi.org/10.1002/jsfa.9466 |
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Journal of the Science of Food and Agriculture |
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99 |
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5 |
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2641 |
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2645 |
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1810431826228936704 |