基于高压结合酶法制备的低敏虾制品体内体外致敏性检测的差异性分析

采用高压结合酶法处理凡纳滨对虾的虾蛋白、虾仁泥(虾肉)和虾仁,获得3种低敏虾制品;采用酶联免疫吸附法检测3种虾制品的过敏原性,采用全身性过敏反应(过敏性休克模型)及致敏离体回肠平滑肌收缩实验(Schultz-Dale反应)检测3种低敏虾制品的致敏性,以分析体内与体外方法检测虾制品致敏性的差异性。结果显示:处理前的3种原料过敏原性极高。经高压结合酶法处理的虾蛋白、虾肉和虾仁的过敏原性分别消减97.0%、94.1%和94.5%。而在动物实验中(以磷酸缓冲盐溶液为阴性对照,未处理虾蛋白为阳性对照,以豚鼠为受试对象),未处理虾蛋白组的过敏反应发生率与死亡率均为100%,3种低敏虾制品的过敏反应发生率也为100%,死亡率分别为16.7%(虾蛋白产物)、50%(虾肉产物)和83.3%(虾仁产物)。致敏的离体回肠受到3种低敏虾制品攻击后,收缩力也明显增强,分别达到376.9%、766.2%和1 004.4%。上述结果表明,对高压结合酶法制备的低敏虾制品过敏原性的检测,体外检测与体内检测存在一定的差异性。

WANG Lijuan,HU Zhihe,WANG Xingxuan,LI Zhiqing,LUO Mengjuan,PAN Zimeng,YANG Yujuan,LIU Lifeng,LIU Li

Three low-allergenic shrimp products, shrimp protein, crushed shelled shrimp and whole shelled shrimp were prepared by combined high pressure and enzymatic treatment. The allergenicity of these products was measured in vitro by enzyme linked immunosorbent assay (ELISA), and by systemic anaphylaxis test in guinea pigs and Schultz-Dale test. A comparative analysis was made between the in vitro and in vivo results. The results showed that the three untreated shrimp samples had extremely high allergenicity, which was reduced by 97.0%, 94.1% and 94.5%, respectively after the combined treatment. In the animal tests using phosphate buffer saline as the negative control and untreated shrimp protein as the positive control, both the incidence of anaphylaxis and mortality in the untreated shrimp protein group were 100%. The incidence of anaphylaxis in the three low-allergenic shrimp products was likewise 100%, and the mortality of shrimp protein product, mashed shrimp product and shrimp meat product were 16.7%, 50% and 83.3%, respectively. The contractility of sensitized isolated ileum challenged by low-allergenic shrimp protein, mashed shrimp and shrimp meat products were increased by 376.9%, 766.2% and 1 004.4%, respectively compared with their untreated counterparts. According to these results, there were differences between in vitro experiments and animal experiments for allergenicity evaluation of low-allergenic shrimp products prepared by combined high-pressure and enzymatic treatment.