https://doi.org/10.4081/ijfs.2025.13551
Set-up and validation of end-point polymerase chain reaction for the detection of edible insect DNA – Acheta domesticus and Tenebrio molitor – in food products
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Published: 14 April 2025
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).
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Böhme K, Calo-Mata P, Barros-Velázquez J, Ortea I, 2019. Review of recent DNA-based methods for main food-authentication topics. J Agric Food Chem 67:3854-64.
Conway A, Jaiswal S, Jaiswal AK, 2024. The potential of edible insects as a safe, palatable, and sustainable food source in the European Union. Foods 13:387.
Debode F, Marien A, Gérard A, Francis F, Fumière O, Berben G, 2017. Development of real-time PCR tests for the detection of Tenebrio molitor in food and feed. Food Addit Contam Part A Chem Anal Control Expo Risk Assess 34:1421-6.
EFSA Scientific Committee, 2015. Risk profile related to production and consumption of insects as food and feed. EFSA J 13:4257.
European Commission, 2017. Commission Implementing Regulation (EU) 2017/2470 of 20 December 2017 establishing the Union list of novel foods in accordance with Regulation (EU) 2015/2283 of the European Parliament and of the Council on novel foods. In: Official Journal L 351/72, 30/12/2017.
European Parliament, Council of the European Union, 2015. Regulation (EU) 2015/2283 of the European Parliament and of the Council of 25 November 2015 on novel foods, amending Regulation (EU) No 1169/2011 of the European Parliament and of the Council and repealing Regulation (EC) No 258/97 of the European Parliament and of the Council and Commission Regulation (EC) No 1852/2001. In: Official Journal, L 327/1, 11/12/2015.
FAO, 2009. How to feed the world in 2050. Available from: http://www.fao.org/fileadmin/templates/wsfs/docs/expert_paper/How_to_Feed_the_World_in_2050.pdf. Accessed on: 13/11/2024.
FAO, 2013. Edible insects – future prospects for food and feed security. Available from: https://www.fao.org/fsnforum/resources/reports-and-briefs/edible-insects-future-prospects-food-and-feed-security. Accessed on: 18/12/2024.
Gałęcki R, Bakuła T, Gołaszewski J, 2023. Foodborne diseases in the edible insect industry in Europe—new challenges and old problems. Foods 12:770.
Geller J, Meyer C, Parker M, Hawk H, 2013. Redesign of PCR primers for mitochondrial cytochrome c oxidase subunit I for marine invertebrates and application in all-taxa biotic surveys. Mol Ecol Resour 13:851-61.
Giusti A, Malloggi C, Magagna G, Filipello V, Armani A, 2024. Is the metabarcoding ripe enough to be applied to the authentication of foodstuff of animal origin? A systematic review. Compr Rev Food Sci Food Saf 23:e13256.
Haiminen N, Edlund S, Chambliss D, Kunitomi M, Weimer BC, Ganesan B, Baker R, Markwell P, Davis M, Huang BC, Kong N, Prill RJ, Marlowe CH, Quintanar A, Pierre S, Dubois G, Kaufman JH, Parida L, Beck KL, 2019. Food authentication from shotgun sequencing reads with an application on high protein powders. NPJ Sci Food 3:24.
Hillinger S, Saeckler J, Domig KJ, Dobrovolny S, Hochegger R, 2023. Development of a DNA metabarcoding method for the identification of insects in food. Foods 12:1086.
Kim TK, Yong HI, Kim YB, Kim HW, Choi YS, 2019. Edible insects as a protein source: a review of public perception, processing technology, and research trends. Food Sci Anim Resour 39:521-40.
Nadeau L, Nadeau I, Franklin F, Dunkel F, 2015. The potential for entomophagy to address undernutrition. Ecol Food Nutr 54:200-8.
Natonek-Wiśniewska M, Krzyścin P, Koseniuk A, 2022. Qualitative and quantitative detection of mealworm DNA in raw and commercial food products using real-time PCR. Genes 13:1400.
Oonincx DGAB, de Boer IJM, 2012. Environmental impact of the production of mealworms as a protein source for humans – a life cycle assessment. PLoS One 7:e51145.
Orsi L, Voege LL, Stranieri S, 2019. Eating edible insects as sustainable food? Exploring the determinants of consumer acceptance in Germany. Food Res Int 125:108573.
Smetana S, Bhatia A, Batta U, Mouhrim N, Tonda A, 2023. Environmental impact potential of insect production chains for food and feed in Europe. Anim Front 13:112-20.
Tramuta C, Gallina S, Bellio A, Bianchi DM, Chiesa F, Rubiola S, Romano A, Decastelli L, 2018. A set of multiplex polymerase chain reactions for genomic detection of nine edible insect species in foods. J. Insect Sci 18:3.
Turck D, Bohn T, Castenmiller J, De Henauw S, Hirsch‐Ernst KI, Maciuk A, Mangelsdorf I, McArdle HJ, Naska A, Pentieva K, Siani A, Thies F, Tsabouri S, Vinceti M, Aguilera‐Gómez M, Cubadda F, Frenzel T, Heinonen M, Maradona MP, Neuhäuser‐Berthold M, Siskos A, Poulsen M, Schlatter JR, van Loveren H, Azzollini D, Knutsen HK, 2024. Safety of Acheta domesticus powder as a Novel food pursuant to Regulation (EU) 2015/2283. EFSA J 22:e8919.
Velasco, A., Ramilo-Fernández, G., Sotelo, C.G., 2020. A real-time PCR method for the authentication of common cuttlefish (Sepia officinalis) in food products. Foods 9:286.
Yazici GN, Ozer MS, 2021. Using edible insects in the production of cookies, biscuits, and crackers: a review. Biol Life Sci Forum 6:80.
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