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Title: Evaluation of a Real-Time Polymerase Chain Reaction (PCR) Assay for Detection of Anisakis simplex Parasite as a Food-Borne Allergen Source in Seafood Products
Authors: Lopez, Itziar; Angel Pardo, Miguel
Citation: JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, 2010, 58, 1469-1477
Abstract: Anisakis simplex has been recognized as an important cause of disease in humans and as a foodborne allergen source. Actually, this food-borne parasite was recently identified as an emerging food safety risk. An A. simplex-specific primer-probe system based on a real-time polymerase chain reaction (PCR) detection assay has been successfully optimized and validated with seafood samples. In addition, a DNA extraction procedure has been optimized to detect the presence of the nematode in food samples. The assay is a very reliable, specific, and sensitive methodology to detect the presence of traces of this parasite in seafood products, including highly processed samples. As a result, 13 sequences of cytochrome c oxidase 11 gene were obtained and scrutinized to calculate intra- and interspecific variabilities of 0 and 35-67\%, respectively. Finally, an efficiency of 2.07 +/- 0.14 of the assay was calculated, and a limit of detection of 40 ppm parasite in 25 g of sample was also optimized. Actually, the presence of this parasite in several seafood products has been demonstrated, enforcing the necessity of a design for a good manufacturing practice protocol for the processing industry to minimize the presence of this parasite as a food-borne allergen source in seafood products.
Keywords: Anisakis simplex; real-time PCR; safety; allergens; seafood; MEAT SPECIES IDENTIFICATION; SANDWICH-ELISA; DNA; EXTRACTION; QUANTIFICATION; CONTAMINATION; RESISTANT; NEMATODES; WHEAT; HEAT
Issue Date: 2010
Publisher: AMER CHEMICAL SOC
Type: Article
Language: English
DOI: 10.1021/jf903492f
URI: http://dspace.azti.es/handle/24689/747
ISSN: 0021-8561
E-ISSN: 1520-5118
Funder: Agriculture and Fisheries Department of the Basque Government
Appears in Publication types:Artículos científicos



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