Detection of allergen mango components by using droplet digital PCR technology

ObjectiveA droplet digital PCR method for detect allergen mango components was established in this study.MethodsFirstly, the chloroplast trnH-psbA gene was selected as the target gene for detect allergen mango components. Secondly, specific primers and probe were designed. Finally, the reaction conditions of droplet digital PCR was optimized to verify the sensitivity and stability of the method.ResultsThis method had been applied to detect allergen mango components in 10 batches of food samples. The results of establishment of droplet digital PCR method was showed good specificity and not cross-react with 10 kinds of other fruits except mango. Under the optimal ddPCR reaction conditions, which were to have the concentration of forward and reverse primers and probe set as 200, 200 and 100 nmol/L, respectively, and to have annealing temperature set as 52 ℃, the detection sensitivity of nucleic acid level reached 5 pg/μL, and the detection sensitivity of mango content reached 5 mg/kg, the coefficient of variation of the method was less than 25%. The test results of 10 batches of food samples applied with the method were consistent with the food labels.ConclusionThis method has the advantages of being fast, specific, and highly sensitive, making it suitable for the detection of allergenic mango components.