Qualitative and Quantitative Detection of Mealworm DNA in Raw and Commercial Food Products Using Real-Time PCR

Considering food safety and an increasing public awareness of the ingredients, production process and origin of foods, the application of insects as food requires the development of tests for the reliable identification of their presence. The aim of the study was (1) the determination of appropriate modifications of the selected method for isolating the DNA of two life stages of mealworm, i.e., larva and adult, from commercial food products; (2) the determination of the method parameters for the qualitative and quantitative analysis of mealworm contents based on the detection of a species-specific mitochondrial DNA fragment, using real-time PCR; (3) the application of a method to test the commercial food products of mealworm. A total of nine species of adult insect were investigated (field cricket, Dubia cockroach, Madagascar cockroach, banded cricket, migratory locust, yellow mealworm, superworm, house fly and lacewing), theirlarvaes (yellow mealworms and superworms) and thirteen commercial food products (dried whole insects, powder and granules) representing various insect species and origins which were purchased from the European market. The obtained results showed that the efficiency of the modification of the DNA extraction method is dependent on the life stage of the mealworm. We proved the high sensitivity of the test, with the range of the method being 0.1-100%; we also proved the biological specificity in this range, and the linearity. The linearity of the test was also statistically verified using the Fisher-Snedecor test. One-way variance analysis showed statistically significant differences between the c_T values of the two mealworm life stages studied, and similarly, between the threshold cycle (c_T) values of adult forms. In contrast, for the inside group of mealworm larvae, there was no significant difference observed between the results of the c_T values. The test is effective for processed food products and may be used to monitor food. The research proved the suitability of the applied method for the analysis of samples that are commercially available as food for exotic animals. The hereby-developed method is based on widely used laboratory techniques, and does not require any additional investment in equipment. The availabilityof such a methodallows for the verification of the accuracy of the declared species component of the food products.