Filling Analytical Gaps in Allergen DetectionReal-Time PCR for the Detection of Commercially Relevant Cephalopods and Gastropods in Food

Mollusks
belong to the group of shellfish, which are considered
to be among the elicitors of severe food allergies worldwide. In recent
years, numerous PCR detection methods have been developed for other
shellfish such as crustaceans. However, cephalopods and gastropods
were not considered in the development of these shellfish detection
systems. In this study, we have developed highly specific real-time
PCR methods for the comprehensive detection of all commercially relevant
cephalopod species and the gastropod families Helicidae, Buccinidae,
and Muricidae in food matrices. In total, we cross-tested over 100
animal and plant species to show the specificity of our systems. The
limit of detection (LOD12) was set at 1 pg of cephalopod
and gastropod DNA or 10 ppm (mg/kg) spiked in a vegetarian food product.
The robustness of the protocol was confirmed by testing multiple parameters
while cooking and autoclaving of samples ensured the practical applicability
of the systems.