Abstract:Objective To establish a detection method for egg components in food using microfluidic digital polymerase chain reaction (PCR) technology.Methods Specific primers and fluorescent probes are designed based on the mitochondrial gene nicotinamide adenine dinucleotide dehydrogenase subunit 1 (ND1). Then, the reaction system and amplification conditions are optimized, and specificity and sensitivity are evaluated before being applied to actual food detection.Results The established method can specifically detect egg components, while no cross-reactivity is observed with other common poultry egg components, such as duck and goose eggs. The optimal annealing temperature is 56 ℃, and the optimal concentrations of primers and probes are 800 nmol/L and 200 nmol/L, respectively. Under these conditions, the limit of detection (LOD) for microfluidic digital PCR is 5.30 copies/μL. Furthermore, the method is applied to detect 20 batches of food samples, yielding consistent results with the sample labels.Conclusion Characterized by anti-interference, high sensitivity, rapidity, and simplicity, the established microfluidic digital PCR method can be applied to detect trace egg components in food.