Abstract
The consumption of insects has increased in western countries, raising concerns about their potential to induce food allergic reactions in sensitized/allergic individuals. This work intended to develop a real-time PCR approach for the detection/quantification of yellow mealworm (Tenebrio molitor) as a potential allergenic food in complex matrices. For this purpose, reference mixtures simulating the production of pork sausages and wheat biscuits containing known amounts of mealworm were used. Real-time PCR with TaqMan probe targeting the cytochrome b gene of T. molitor was able to detect up to 2 fg of insect DNA, and 1.0 and 0.1 mg/kg of mealworm flour in autoclaved sausages and baked biscuits, respectively. Generally, the method showed acceptable analytical performance parameters, confirming its suitability/applicability for a wide range of foods. However, real-time PCR data showed significant differences among food matrix and processing, highlighting the importance of using appropriate calibration models for quantitative analysis. Finally, the real-time PCR approach was successfully validated with blind mixtures and applied to commercial samples, demonstrating its efficacy and reliability in the quantification of mealworm in processed foodstuffs.