Author(s): ZHANG Xia, ZHAO Liang-juan, GAO Lin, WEN Hua-wei, WANG Fei-xue, DONG Yan, LIU Jue-mang, LIU Yin, ZHENG Wen-jie
Pages: 148-151
Year: 2016 Issue:  9
Journal: Food Research and Development
Keyword:  peanut allergen; proximity ligation assay; affinity probe; magnetic particles;
Abstract: To establish a new method with high-sensitivity for detection of peanut allergen-proximity ligation assay. The proximity ligation assay compound consists of monoclonal antibody from mouse to peanut allergen, polyclonal antibody from rabbit to peanut allergen, secondary antibody goat anti-mouse, secondary antibody goat anti-rabbit, DNA nucleotide probe 1 and 2, DNA connector. Three kinds of peanut allergen reference standards and ten samples without peanut allergen including hazelnut and apricot seed references were evaluat-ed the specificity of the proximity ligation assay. The dilution of the peanut allergen reference standards were used to evaluate the sensitivity of the assay. The assay established in present study was highly specific and sen-sitive. The limit of detection was 0.01 ng/mL for the peanut allergen reference standards, which was 1 000 times better than the ELISA method. The method can detect the invisible peanut allergen objects using traditional methods. It is very useful for detection of peanut allergen in the actual detection work.