Development and validation of a novel real-time PCR protocol for the detection of buffalo worm (Alphitobius diaperinus) in food

The year 2021 represented a benchmark for the insect feed and food sector: while new species were authorized for the first time for human consumption, for other insects the authorization as feed for aquaculture was extended to poultry and swine. While these statutory provisions represent a huge incentive for the sector, consumers’ safety needs to be safeguarded. This includes as well the setting up of robust laboratory methods for the authentication of these novel products. We present the development and validation of a real-time PCR-based method for the identification of the buffalo worm Alphitobius diaperinus in foods. The specificity of the system was confirmed with a broad range of plant and animal species. The technical limit of detection with pure A. diaperinus DNA was lower than a single genome copy. In food, we observed a sensitivity limit ranging from 1 to 20 ppm, depending on the matrix analyzed and on the degree and the nature of food processing. Practical applicability of the proposed system was tested on 31 commercial foods declaring the presence of A. diaperinus as well as of other insects. Based on the validation carried out, the tested system is suitable for routine tests, enabling the sensitive and reliable detection of the buffalo worm A. diaperinus in complex and processed foods.