1.Preparation and identification of protein chip for detecting components of peach allergen
le Kang ZHAI ; xiong Jun ZHOU ; xin Xin LI ; Hui CHEN ; Jia YIN ; Wei HE ; min Jian ZHANG
Basic & Clinical Medicine 2017;37(11):1507-1512
Objective To prepare a protein chip for the detection of serum antibodies against several peach allergen components, and to provide a rapid and reliable method for clinical diagnosis of peach allergic disease .Methods The coding sequences of six key peach components Pru p 1, Pru p 2, Pru p 3, Pru p 4, Pru p 7 and Pru p G were inserted into pGAPZαA yeast expression vector.Then the vectors were digested by endonuclease Avr Ⅱ and electroporated into yeast SMD1168 for protein expression.After peach proteins were purified, the protein chip was prepared and used to detect the antibodies against peach allergen components in serum samples from 41 suspected patients.The sensitivity and specificity of the protein chip were verified by comparison with ImmunoCAP method .Res ults The protein chip was prepared with these proteins for detecting IgE antibodies in serum samples against these four peach allergen components.The results showed that the sensitivity of the protein chip to Pru p 1, Pru p 3 and Pru p 4 was 40%, 100%and 100%, and the specificity was 100%, 50%and 100%respectively.The total sensitivity and specificity was 86%and 96%respectively.Six serum samples from peach allergy patients were identified Pru p7 antibody positive by the protein chip.Co nclusions The sensitivity and specificity of the protein chip is comparable to ImmunoCAP.It needs less serum and so to be a potential method for the clinical diagnosis of peach allergenic disease after optimization.