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.
abstract: Anisakis simplex has been recognized as an important cause of disease in humans and as a food-borne 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 II 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.
authors.affiliation: Food Research Division, Fisheries and Food Technological Institute, Parque Tecnologico de Bizkaia, Astondo Bidea-Edificio 609, E-48160 Derio, Bizkaia, Spain.
authors.forename: Itziar
authors.initials: I
authors.lastname: Lopez
authors.forename: Miguel Angel
authors.initials: MA
authors.lastname: Pardo
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017163:Anisakis / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D000947:Antigens, Helminth / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D017931:DNA Primers / Q000235:genetics; D003576:Electron Transport Complex IV / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D005399:Fishes / Q000469:parasitology*; D057230:Limit of Detection; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D017747:Seafood / Q000032:analysis* / Q000469:parasitology; D016415:Sequence Alignment
chemical_list: D000947:Antigens, Helminth; D017931:DNA Primers; C117108:cytochrome C oxidase subunit II; D003576:Electron Transport Complex IV