Department of Chemistry, Biochemistry and Food Microbiology, Food Research Institute Prague, Radiová 1285/7, Prague 10 102 00, Czech Republic.
Department of Biochemistry and Microbiology, University of Chemistry and Technology, Prague, Technická 5, Prague 6 166 28, Czech Republic.
J Agric Food Chem. 2024 Jun 5;72(22):12788-12797. doi: 10.1021/acs.jafc.4c01410. Epub 2024 May 23.
Fish from the pike () 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.
来自鱒属的鱼类因其优质的肉质而在美食界受到重视。然而，它们可能会引起敏感消费者的过敏反应。本工作旨在利用分子生物学技术填补鱒过敏原检测的空白。使用副肌球蛋白基因为标记物，设计了新的、快速且准确的环介导等温扩增（LAMP）和实时聚合酶链式反应（qPCR）检测方法来检测鱒 DNA。通过电泳、SYBR 绿光学检测和实时荧光检测评估 LAMP。后者是最敏感的，可检测到低至 0.78ng 的鱒 DNA；qPCR 的检测限为 0.1ng。LAMP 分析需要 20-70 分钟，明显快于 qPCR。本研究提供了新鲜和加工样品中副肌球蛋白基因的可靠检测和定量，进一步强调了使用副肌球蛋白基因进行食品产品认证和消费者保护的多功能性，通过独立于所使用的组织类型或食品加工方法的精细过敏原风险评估来实现。
