Abstract
The analysis of food for allergens is an inherent part of consumer protection against health risks. Before application, these methods require validation by collaborative studies to ensure comparable results between laboratories. In this article, we present results of collaborative trial studies for validating RT-PCR methods for the detection and quantification of two food allergens, fish and peanut. Reference materials of different food matrices, spiked with fish and peanut, respectively, were analysed. Defined food matrix DNA standards (fish, peanut) as well as copy number based DNA solutions (fish) were applied for calibration and quantification. A limit of detection of approximately 10–20 mg/kg dried fish (i.e. approx. 50 mg fresh weight/kg) could be confirmed within a collaborative trial performed by 13 participating labs. Trueness and precision data were comparable to former collaborative studies suggesting that quantification is possible, if the fish species is known and appropriate calibration material is available. For peanut, we used a multicopy target-sequence, which allows a very sensitive detection of this allergen. Within the collaborative trial of 14 laboratories, a peanut concentration of 0.5 mg/kg (i.e. approx. 0.1–0.2 mg peanut protein/kg) could be detected reliably in a processed cookie matrix. The method revealed acceptable trueness and good precision data and is therefore suitable for quantification of the allergenic ingredient peanut. Both methods will be included into the Official Collection of Methods of Analysis in Germany in 2019 as a national standard method.
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Acknowledgements
We thank the members of the working group “Food Allergens” within the scope of the official method collection according to §64 of the German Food and Feed Code (LFGB) for their participation in the study and their fruitful comments: Daniela Bartsch, Janine Beutlich, Ulrich Busch, Gabriele Engler-Blum, Thomas Gerhardt, Ilka Haase, Rupert Hochegger, Thomas Holzhauser, Sandra Kerbach, Kirsten Kerkhoff, Ingo Krujatz, Marion Mägdefrau, Edith Müller, Gabriele Näumann, Katrin Neumann, Angelika Paschke-Kratzin, Gisela Ohrt, Bert Pöpping, Petra Reihl, Martin Röder, Anke Rullmann, Annette Sauer, Fredi Schwägele, Rolf Steinmüller and Jutta Zagon.
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Waiblinger, HU., Boernsen, B., Geppert, C. et al. Collaborative trial validation of RT-PCR methods for the detection and quantification of the allergenic foods fish and peanut. J Consum Prot Food Saf 14, 301–311 (2019). https://doi.org/10.1007/s00003-019-01235-3
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DOI: https://doi.org/10.1007/s00003-019-01235-3