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

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Journal of Ocean University of China 2022年第2期21卷 421-429页

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 Key Laboratory of Aquatic Biotechnology Ministry of EducationNingbo UniversityNingbo 315211China Collaborative Innovation Center for Zhejiang Marine High-Efficiency and Healthy Aquaculture Ningbo UniversityNingbo 315211China The Affiliated Hospital of Medical School of Ningbo University Ningbo 315020China

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.

关键词： Portunus trituberculatus potential allergens food anaphylaxis ELISA reagent

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