Abstract
Food allergies represent a clear threat to the general health and wellbeing of those affected which place increasing pressure on food producers and regulatory authorities. Current analytical techniques typically find difficulties distinguishing between closely related Prunus species which include almond (Prunus dulcis), an EU listed allergenic species. This study describes a proof of principle real-time PCR approach utilising DNA melt analyses that targets the internal transcribed spacer (ITS) sequence to differentiate between a panel of Prunus test species. The method was successfully applied to the characterisation of a commercial paprika sample suspected of having being adulterated with almond, referred to the UK Government Chemist in 2015 in its advisory capacity. Subject to further validation work, the method appears to specifically amplify Prunus species and is capable of discrimination based on the resultant melt profiles. The developed method provides analysts with a simple and broad molecular tool to identify common Prunus species for food authenticity and allergen testing purposes. Initial development work demonstrates a promising approach with the potential to improve discrimination between Prunus species not easily resolved by routine analytical methods.
Highlights
Food allergies, characterised by adverse immunologic (IgE and non-IgE mediated) reactions to food, have po-How to cite this paper: Nixon, G., Hall, L., Wilkes, T., Walker, M. and Burns, M. (2016) Novel Approach to the Rapid Differentiation of Common Prunus Allergen Species by polymerase chain reaction (PCR) Product Melt Analysis
GenBank® [25] Entrez Nucleotide sequence database searches and ClustalW [28] alignments showed conserved and variable regions with related Prunus species which supported the application of the assay to species differentiation by melt analysis
A preliminary evaluation study was conducted that challenged the DNA-binding dye-based real-time PCR assay with a panel of commonly found Prunus species, cumin and paprika control materials, and a paprika-based commercial sample referred to the UK Government Chemist for further analysis
Summary
Food allergies, characterised by adverse immunologic (IgE and non-IgE mediated) reactions to food, have po-. (2016) Novel Approach to the Rapid Differentiation of Common Prunus Allergen Species by PCR Product Melt Analysis. Prunus is a large genus and includes trees and shrubs, such as almond, plums, cherries, peaches, nectarines, apricots and mahaleb Several of these important species contain allergenic proteins [19] only almond is listed in Annex II of EU Regulation 1169/2011, obliging disclosure when used in food [10]. We report the development and demonstrate proof of principle for a simple real-time PCR screening approach utilising melting analysis to discriminate between closely related Prunus species. The PCR screening approach reported will augment the capabilities of both Official Control and food industry laboratories by providing a means to differentiate between common Prunus species
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