虾蟹类食物过敏者血清的筛选与鉴定  

Screening and identification of human serum allergic to crabs and shrimps

作　　者：沈苑[1] 饶世涛[1] 曹敏杰[1] 李兵[1] 蔡秋凤[1] 刘光明[1] 

机构地区：[1]集美大学生物工程学院,福建省高校水产科学技术与食品安全重点实验室,厦门361021

出　　处：《食品科技》2010年第6期312-317,共6页Food Science and Technology

基　　金：国家自然科学基金项目(30871947);福建省自然科学基金项目(8J0067);集美大学创新团队基金项目(2008A002);2007年度福建省高等学校新世纪优秀人才支持计划项目

摘　　要：近年来食物过敏性疾病呈上升趋势,虾蟹等甲壳类动物是联合国粮农组织公布的八大食物过敏原之一,虾蟹类过敏严重影响过敏人群的身体健康和生活质量。通过纯化虾和蟹过敏蛋白对集美大学新生血清进行ELISA筛选分析,以期获得对虾和蟹过敏的阳性血清。结果通过问卷调查收集到了397份自诉虾蟹过敏者血清,用硫酸铵盐析、等电点沉淀和沸水浴加热的方法从南美白对虾中纯化到了电泳纯的原肌球蛋白,并用兔抗锯缘青蟹原肌球蛋白的Western-blot进行了检测确认;用硫酸铵盐析和High-Q柱层析方法从锯缘青蟹中纯化到了电泳纯的精氨酸激酶,并用鼠抗白虾精氨酸激酶的Western-blot进行了检测确认;然后利用2种纯化蛋白的ELISA分析397份自诉虾蟹过敏者血清,从中筛选获得28份对原肌球蛋白呈阳性反应的血清,21份对精氨酸激酶呈阳性反应的血清,最后采用Dot-blot实验对筛选阳性反应的血清进行了确认。Adverse food reactions in hypersensitive individuals are more and more serious in recent years,and crustaceans such as shrimps and crabs are one of the eight kinds of foods which are easily cause allergy reported by FAO.Crustacean hypersensitivity reactions affect the allergy crowds＇ health and life quality seriously.For this reason,it is necessary for us to do some research on the allergens of shrimp and crab.In this paper,using ELISA to screen and analyses the Jimei university students＇ sera by the purified allergic proteins from shrimp and crab so as to obtain the shrimp and crab allergy positive serum.The result show us that we have obtained 397 cases private prosecution allergic sera through a questionnaire survey,then use the method such as ammonium sulfate salting out,isoelectric point precipitation and over the High-Q column to pure the gel electrophoresis purification of tropomyosin and arginine kinase which are confirmed by Western-blotting.And then using the purified protein to analysis the sera by ELISA method and have screened 28 sera which have a positive reaction to tropomyosin and 21 sera which have a positive reaction to arginine kinase from the 397 cases sera,and Dot-blot experiments confirmed positive.

关 键 词：南美白对虾 原肌球蛋白 锯缘青蟹 精氨酸激酶 过敏原 纯化 免疫检测 

分 类 号：R446.6[医药卫生—诊断学]