An improved duplex real-time PCR method for the systematic detection of commercially relevant crustaceans in food

Crustaceans are known food allergens that can cause severe reactions for allergic patients. Hitherto, methods to detect commercial relevant crustacean species often consist of many PCR systems. In this study, we developed a highly specific duplex PCR (polymerase chain reaction) method for the detection of most commercially relevant crustaceans in food. The nuclear 18S rRNA and the mitochondrial 16S rRNA genes were selected for the detection of the suborders Pleocyemata and Dendrobranchiata, respectively. Both suborders belong to the order Decapoda, which comprises about 96% of all crustaceans consumed as food. We cross-tested over 100 different plant and animal species, including 25 species of crustaceans and eight species of farmed insects. Eight different food products were tested, to ensure practical applicability. The limit of detection (LOD12) is 1 pg of crustacean DNA or 10 ppm (mg/kg) of crustacean sample in food matrix. Finally, a robustness test confirmed the practical use of the protocol.

Automated summary

In this study, a highly specific duplex PCR method was developed for the detection of most commercially relevant crustaceans in food. The limit of detection was 1 pg of crustacean DNA or 10 ppm of crustacean sample. Cross-testing with various plant and animal species ensured practical applicability, and a robustness test confirmed the practical use of the protocol.