Čermáková Eliška, Svoboda Pavel, Ovesná Jaroslava, Vašek Jakub, Demnerová Kateřina, Zdeňková Kamila
Department of Biochemistry and Microbiology, Faculty of Food and Biochemical Technology, University of Chemistry and Technology, Technická 5, 166 28 Prague, Czech Republic.
Department of Chemistry, Biochemistry and Food Microbiology, Food Research Institute Prague, Radiová 1285/7, 102 31 Prague, Czech Republic.
Foods. 2024 May 7;13(10):1432. doi: 10.3390/foods13101432.
This paper discusses the development of rapid, reliable, and accurate polymerase chain reaction (PCR) assays for detecting opium poppy ( L.) in food. Endpoint, quantitative, and digital PCRs were compared based on the amplification of a newly developed DNA marker targeting the NADPH-dependent codeinone reductase (COR) gene. Designed assays were shown to be highly specific and sensitive in discriminating opium poppy from other plant species, even in heat-treated and food samples. Digital PCR was the most sensitive, with a detection limit of up to 5 copies, i.e., approximately 14 pg of target DNA per reaction. Quantitative and digital PCR further allowed the quantification of opium poppy in up to 1.5 ng and 42 pg (15 copies) of target DNA in a sample, respectively. In addition, two duplex PCRs have been developed for the simultaneous detection of opium poppy DNA and representatives of (i) the Papaveraceae family or (ii) the Plantae kingdom. Finally, all designed assays were successfully applied for analysis of 15 commercial foodstuffs; two were suspected of being adulterated. The study results have an important impact on addressing food fraud and ensuring the safety and authenticity of food products. Beyond food adulteration, the study may also have significant implications for forensics and law enforcement.
本文讨论了用于检测食品中罂粟（Papaver somniferum L.）的快速、可靠且准确的聚合酶链反应（PCR）检测方法的开发。基于针对依赖烟酰胺腺嘌呤二核苷酸磷酸（NADPH）的可待因酮还原酶（COR）基因新开发的DNA标记物的扩增，对终点PCR、定量PCR和数字PCR进行了比较。所设计的检测方法在区分罂粟与其他植物物种时显示出高度的特异性和敏感性，即使在经过热处理的样品和食品样品中也是如此。数字PCR最为灵敏，检测限高达5个拷贝，即每个反应约14 pg的靶DNA。定量PCR和数字PCR还分别能够对样品中高达1.5 ng和42 pg（15个拷贝）的靶DNA中的罂粟进行定量。此外，还开发了两种双重PCR，用于同时检测罂粟DNA以及（i）罂粟科或（ii）植物界的代表物种。最后，所有设计的检测方法都成功应用于15种商业食品的分析；其中两种被怀疑掺假。研究结果对解决食品欺诈以及确保食品产品的安全性和真实性具有重要影响。除了食品掺假问题，该研究对法医学和执法也可能具有重大意义。
