IgE Reactivity of Potential Allergens from the Swimming Crab Portunus trituberculatus and the Research of ELISA Reagent for Detecting Crab Food Anaphylaxis

作     者：QIU Xueni ZHOU Qiongyan ZHU Xiaoxia LIN Wei MU Changkao LI Ronghua YE Yangfang SONG Weiwei SHI Ce LIU Lei WANG Huan WANG Chunlin XU Suling QIU Xueni;ZHOU Qiongyan;ZHU Xiaoxia;LIN Wei;MU Changkao;LI Ronghua;YE Yangfang;SONG Weiwei;SHI Ce;LIU Lei;WANG Huan;WANG Chunlin;XU Suling

作者机构：Key Laboratory of Aquatic BiotechnologyMinistry of EducationNingbo UniversityNingbo 315211China Collaborative Innovation Center for Zhejiang Marine High-Efficiency and Healthy AquacultureNingbo UniversityNingbo 315211China The Affiliated Hospital of Medical School of Ningbo UniversityNingbo 315020China 

出 版 物：《Journal of Ocean University of China》 (中国海洋大学学报（英文版）)

年 卷 期：2022年第21卷第2期

页      面：421-429页

核心收录：

学科分类：0832[工学-食品科学与工程（可授工学、农学学位）] 08[工学] 083204[工学-水产品加工及贮藏工程] 

基　　金：supported by Zhe-jiang Provincial Natural Science Foundation of China(No.LY18H110003) the General Scientific Research Pro-ject of Zhejiang Education Department(No.Y201940887) 

主　　题：Portunus trituberculatus potential allergens food anaphylaxis ELISA reagent 

摘      要：Crustacean is one of the major allergic *** is of great significance to identify more crab allergens and research the detection methods for crab food *** this study,IgE reactivity to three recombinant proteins from Portunus trituberculatus,including tropomyosin(rPtTM),myosin light chain(rPtMLC),and pancreatic lipase(rPtPL),were detected by enzyme-linked im-munosorbent assay(ELISA).The expressions of tropomyosin(TM)in various tissues of *** were detected by Western blot(WB).Furthermore,microplates were coated with rPtTM and the ELISA conditions were *** cut-off value was deter-mined by the receiver operating characteristic(ROC)*** 51 crab-allergic sera,21(41.2%)showed positive IgE to *** 70 crab-allergic sera more frequently recognized rPtPL(9/70;12.9%),followed by rPtMLC(1/70;1.4%).WB results showed that TM was mainly expressed in the muscle,followed by the heart and a small amount in gills and the *** optimal results showed that the coating condition of rPtTM was 50 ng per well with coating for 3 h at 37℃.The optimal blocking condition was 1.2%BSA with blocking for 3 h at 37℃,and the optimal dilution of the second antibody was 1:*** ROC curve showed that the ELISA reagent had high sensitivity(83.52%)and specificity(98.00%)when the cut-off value was *** results indicated that tropomyosin is the major allergen of ***,and myosin light chain and pancreatic lipase are the potential ***-tionally,the ELISA reagent developed with the rPtTM was feasible for laboratory detection of crab anaphylaxis.