Exploiting 16S rRNA gene for the detection and quantification of fish as a potential allergenic food: A comparison of two real-time PCR approaches

• Type: Article in International Scientific Journal
• Date: 2018

Authors

Fernandes, TJR (Author) Other; Joana Costa (Author) FFUP; Beatriz B P P Oliveira (Author) FFUP; Mafra, I (Author) Other

Journal

Title: Food Chemistry

Vol. 245

Pages: 1034-1041

ISSN: 0308-8146

Publisher: Elsevier

Indexing

ISI Web of Knowledge - 5 Citations

Scopus - 11 Citations

Other information

Authenticus ID: P-00N-B69

DOI: 10.1016/j.foodchem.2017.11.068

Abstract (EN): Fish is one of the most common allergenic foods that should be accurately labelled to protect the health of allergic consumers. In this work, two real-time PCR systems based on the EvaGreen dye and a TaqMan probe are proposed and compared. New primers were designed to target the 16S rRNA gene, as a universal maker for fish detection, with fully demonstrated specificity for a wide range of fish species. Both systems showed similar absolute sensitivities, down to 0.01 pg of fish DNA, and adequate real-time PCR performance parameters. The probe system showed higher relative sensitivity and dynamic range (0.0001-50%) than the EvaGreen (0.05-50%). They were both precise, but trueness was compromised at the highest tested level with the EvaGreen assay. Therefore, both systems were successful, although the probe one exhibited the best performance. Its application to verify labelling compliance of foodstuffs suggested a high level of mislabelling and/or fraudulent practices.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 8