Novel Approach to the Rapid Differentiation of Common <i>Prunus</i> Allergen Species by PCR Product Melt Analysis

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.