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- Advanced botanical authentication of honey: Using an ultrasensitive electrochemical genosensor and RT-qPCR for the detection of Castanea sativaPublication . Morais, Stephanie; Pereira, Eduarda; Ferreira, Mariana; Santos, Marlene; Soares, Sónia; Texeira, Ana L.; Domingues, Valentina; Delerue-Matos, Cristina; Barroso, M. Fátima; Santos, MarleneFood fraud is a reoccurring issue for the food industry, with significant public health and economic implications. Honey, a natural ingredient prized for its sweetness and inherent nutritional profile and health benefits, is one of the most frequently adulterated foods found in the international market. This fraudulent act not only damages the reputation of the honey industry but also presents a hazard to the consumers’ health. So, in this study, a disposable electrochemical genosensor was developed to detect Castanea sativa (chestnut tree) DNA in commercial honey samples. For this, a 103 bp C. sativa specific DNA-target oligonucleotide and its complementary probe were selected and designed. The genosensor methodology implied a sandwich hybridization format, for which the complementary sequence was cut into a 22 bp thiolated DNAcapture probe and an 81 bp fluorescein isothiocyanate-labelled DNA-signaling probe. Using chronoamperometric measurements, the enzymatic amplification of the electrochemical signal was obtained in a 0.03 to 1.00 nM concentration range, with a LOD and LOQ of 3.01 and 10.04 pM, respectively. The developed genosensor was able to detect the presence of the chestnut DNA in real chestnut plants and commercial honey samples. These results were then validated real-time quantitative PCR (RT-qPCR). In fact, conventional PCR coupled with gel electrophorese was not able to detect the presence of heather in honey. Therefore, electrochemical genosensors are a promising and cost-effective analytical tool to authenticate the botanical origin of honey, guaranteeing its safety, quality and authenticity.