The Research Of Multiplex PCR Assays To Identify Components Of Hazelnut And Its Allergens In Food

This research focused on the establishment of a multiplex PCR to detect the components of hazelnut and its major allergens. The three components detected by this research were cytosolic phosphoglucose isomerase, Cor a 1 and Cor a 8. Different lysates were used to schizolysis samples. A suitable method was established to extract the DNA from nuts by comparing the results of different extract methods. Corresponding primers were designed by professional softwares Primer3 and DNAstar, based on the three different target proteins. Primers with strong specificity were selected by comparing the results of PCR amplification and BLAST. Further selection was carried out by multiplex PCR in order to determine the three most suitable pairs of primers for triplex-PCR, the results of PCR amplification were also confirmed by sequencing. The PCR amplification productions were analyzed by Agarose Gel Electrophoresis in the Gel Imaging System. The results showed that the methods established in this research performed good specificity, and the detection limit could reach 10 ng/μL. Consequently, it's a suitable detection method for hazelnut and allergens of hazelnut in food.