Fish from the pike (Esox) genus are valued in gastronomy for their superior meat quality. However, they can cause allergic reactions in sensitive consumers. This work aimed to fill the gap in the detection of pike allergens using molecular-biological techniques. New, fast, and accurate loop-mediated isothermal amplification (LAMP) and real-time PCR (qPCR) assays were designed to detect pike DNA using the parvalbumin gene as a marker. LAMP was assessed by electrophoresis, SYBR green optical detection, and real-time fluorescence detection. The latter was the most sensitive, detecting as little as 0.78 ng of pike DNA; the qPCR detection limit was 0.1 ng. The LAMP analysis took 20-70 min, which is significantly faster than qPCR. The study provides reliable detection and quantification of the parvalbumin gene in both fresh and processed samples and further highlights the versatility of the use of the parvalbumin gene for the authentication of food products and consumer protection via refined allergen risk assessment that is independent of the type of tissue or food processing method used.

Department of Biochemistry and Microbiology University of Chemistry and Technology Prague Technická 5 Prague 6 166 28 Czech Republic

Department of Chemistry Biochemistry and Food Microbiology Food Research Institute Prague Radiová 1285 7 Prague 10 102 00 Czech Republic

Institute for Environmental Studies Faculty of Science Charles University Benatska 2 Prague 2 128 01 Czech Republic

Lennard Jones School of Chemical and Physical Sciences Keele University Staffordshire ST5 5BG United Kingdom

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