Detection of peanut allergens by means of new PCR based methods and ELISA

Peanut is one of the most allergenic foods since even low amounts of this ingredient can trigger severe allergic reactions up to life-threatening symptoms. The goal of this thesis was therefore to design and develop new DNA based methods for detection of peanut allergens residues in real foodstuffs. Three real-time PCR assays were developed and applied to analysis of cookies and chocolate model matrices with particular regard to respectively the effect of heat treatment and cocoa content. A comparative study between real-time PCR and protein based kits was performed on hundreds of market samples showing that, although different peanut target molecules are detected, real-time PCR and ELISA offer a comparable performance. Design of new methods also embraced the applicability of innovative chemistries based on PNA probes. Circular dichroism, microarray and light up probe were the technologies studied which showed the potential of PNA in the detection of peanut traces in real food samples.