Development and validation of two real-time PCR-methods for the rapid detection of commercially available mussels and scallops in food

Food allergies against bivalves are one of the most common chronic health conditions in the world. With no existing cure, allergy sufferers must rely on proper labeling of food contaminants. In this study, we developed two bivalve real-time PCR systems for the detection of the commercially relevant species of the two orders Mytilida and Pectinida. We have tested 23 species of bivalve molluscs belonging to the above orders and have cross-tested our systems with over 80 different animals and plants in order to demonstrate the specificity of our systems. The Limit of detection (LOD₁₂) is 1 pg for the Mytilida system and 0.1 pg for the Pectinida system. In food matrices a sensitivity of 10 mg/kg of species per food matrix has been achieved for both systems. In addition, practical application of the protocol was ensured by testing cooked and autoclaved food samples. Finally, the robustness of the system has been confirmed by testing multiple PCR parameters.