甲壳类水产品主要过敏原原肌球蛋白胶体金免疫层析快速检测方法的建立及应用  

Establishment and application of colloidal gold immunochromatographic method for rapid detection of major allergen tropomyosin in crustacean aquatic products

作　　者：高红艳 赵金龙 王雅晴 蒋秋燕[3] 黄玉浩 林洪[1] 李振兴[1] GAO Hongyan;ZHAO Jinlong;WANG Yaqing;JIANG Qiuyan;HUANG Yuhao;LIN Hong;LI Zhenxing(Ocean University of China,College of Food Science and Engineering,Qingdao 266003,China;Qingdao Marine Food Nutrition and Health Innovation Research Institute,Qingdao 266109,China;Shandong Institute of Commerce and Technology,Jinan 250103,China)

机构地区：[1]中国海洋大学食品科学与工程学院,山东青岛266003 [2]青岛海洋食品营养与健康创新研究院,山东青岛266109 [3]山东商业职业技术学院,山东济南250103

出　　处：《水产学报》2022年第7期1269-1281,共13页Journal of Fisheries of China

基　　金：国家自然科学基金(32072338)。

摘　　要：为实现甲壳类水产品主要过敏原的快速检测,实验使用凡纳滨对虾原肌球蛋白(TM)作为检测靶标,构建了胶体金免疫层析检测方法。使用天然TM分别免疫SD大鼠和新西兰大白兔制备多克隆抗体,以40 nm胶体金标记的鼠多抗作为检测抗体,以兔多抗为捕获抗体,基于双抗体夹心原理组装免疫层析试纸条,并应用于市售食物样品的检测。结果显示,组建的免疫层析试纸条可在5 min之内显示结果,视觉检出限为10 ng/mL,对虾、蟹、克氏原螯虾及美洲螯龙虾等甲壳类水产品的特异性高,但与蛤蜊、扇贝存在微弱的交叉反应;在不含甲壳类水产品的市售食品中的加标回收率为81%~126%,准确性良好;试纸条的批内、批间变异系数低于15%,市售实际食物样品的检测结果与食物过敏原标签一致。研究表明,该方法具有高灵敏度与检测特异性,可在多种基质与商业化食品中实现甲壳类过敏原的有效检测,具有较强的实际应用价值。Currently, food allergy has become a food safety issue of global concern, and there is no effective cure at home and abroad. Avoiding the intake of allergens is the best solution to prevent food allergies;therefore, the construction of a rapid and sensitive allergen detection system is of great research significance to ensure the safety of consumers. A colloidal gold immunochromatographic method was established with tropomyosin(TM) of Litopenaeus vannamei as the detection target for the rapid detection of major allergens TM in crustacean aquatic products. SD rats and New Zealand rabbits were immunized with natural TM to prepare polyclonal antibodies.Then, the immunochromatographic test strips were assembled based on the double antibody sandwich principle using 40 nm colloidal gold labeled rat anti-TM IgG as detection antibodies and rabbit anti-TM IgG as capture antibodies for the detection of commercial food samples. The test strip can display the results within 5 min with a 10ng/mL visual detection limit. The developed immunochromatographic test strips showed high specificity to crustacean, such as shrimp, crab, Procambarus clarkii and Homarus americanus, although there is a weak cross reaction with clams and scallops. The recoveries of standard addition in commercial food products without crustaceans were 81%-126%, indicating the method’s high accuracy. The intra-and inter-batch coefficient of variation of the test strip was less than 15%. Additionally, the test results of the actual food samples were consistent with the food allergen label. The results show that the developed method possesses sufficient sensitivity and specificity. Therefore, it can effectively detect crustacean residues in a variety of matrices and commercial food samples and has strong practical application value.

关 键 词：凡纳滨对虾 甲壳类 原肌球蛋白 胶体金免疫层析 过敏原检测 

分 类 号：TS254.7[轻工技术与工程—水产品加工及贮藏工程]