Preliminary Study on PCR Detection Method for Food Allergen Oyster

Objective:A PCR based method combined with melting curve analysis and site-specific Tm and Ct analysis was established to detect oyster as allergen in different types of food. Methods:According to the DNA sequences of oyster mitochondria from NCBI database, oyster-specific primers for both conventional PCR and SYBR Green I RT-qPCR were designed by DNA star software. 10 samples were confirmed to contain oyster (positive) and 20 samples were negative in the oyster presentation. Some samples made of the oyster were used in our experiments as positive controls. Results:The minimum detectable proportion of oyster component in food samples was 0.1% (w/w), with a 0.01 ng/μL detection limit of qPCR and a melt curve peak at Tm 80.08℃ by this methods. Conclusion:Compared with the existing methods for oyster detection in food industry, the qPCR based protocol established here was step-wise, cost-effective with higher specificity and sensitivity. In this study, the detection efficiency of oyster components in all collected samples was 100%, which indicated this method would be widely applied for quick qualification of oyster component in food industry.