Set-up and validation of end-point polymerase chain reaction for the detection of edible insect DNA – <i>Acheta domesticus</i> and <i>Tenebrio molitor</i> – in food products

Insects offer an alternative protein supply for both animal and human consumption. Recently, companies in Europe have begun insect production for use in food and animal feed. Following Regulation (EU) No. 2015/2283, edible insects are classified under the novel food category and can be marketed and consumed within the European Union. As a result, methods of authentication are necessary to verify the compliance of these insect-based products. In this study, two end-point polymerase chain reaction assays for the detection of the 16S rRNA gene of Acheta domesticus and Tenebrio molitor were fine-tuned and validated. The primer specificity was tested on insect samples, other animals, and plants, and the gene was amplified only in target samples. The limit of detection was evaluated using different food matrices contaminated with decreasing concentrations of insect powder and was below 0.05% (w/w).