伏马毒素和氯霉素免疫亲和检测柱的研究

发布时间：2018-06-30 05:20

本文选题：伏马毒素 + 氯霉素　；参考：《天津科技大学》2012年硕士论文

【摘要】：论文采用一种免疫亲和凝胶柱对食品中伏马毒素B1(FB1)和氯霉素(CAP)残留的检测进行了系统研究,用溴化氢活化的琼脂糖凝胶偶联伏马毒素抗体或氯霉素抗体制备其分别的抗体胶；偶联辣根过氧化氢酶(HRP)抗体制备其HRP胶；将溴化氢活化的琼脂糖凝胶封闭制备封闭胶。封闭胶与抗体胶混合制备成检测层,与HRP胶混合制备成对照层,装入检测柱,并对影响实验的各个因素(离子强度,pH值等)进行了优化,建立了一种省时、灵敏度高、可视化的检测方法,可以定性半定量的检测食品中的伏马毒素和氯霉素。对于伏马毒素B1免疫亲和凝胶检测柱：封闭胶与抗体胶以1：10比例混合可以达到良好的视觉显色结果,封闭胶与抗体胶1：90混合制备其对照层,其FB1-HRP以1：8000稀释用于凝胶检测柱的检测,建立伏马毒素免疫亲和凝胶检测柱的最低限量检测限(LOD)为40μg/L。对啤酒、白酒、玉米粉、谷物牛奶中伏马毒素残留进行了检测：啤酒和白酒直接稀释5倍；玉米粉加入PBS提取离心后稀释5倍；谷物牛奶离心取上层溶液稀释10倍。对于氯霉素免疫亲和凝胶检测柱：封闭胶与抗体胶以1：20比例混合是可以达到良好的视觉显色结果,封闭胶与抗体胶1：100混合制备其对照层,其CAP-HRP以1：20万稀释用于凝胶检测柱的检测,建立氯霉素免疫亲和凝胶检测柱的LOD为1μg/L。对牛奶、乳粉、鱼塘水、蜂蜜中氯霉素的残留进行了检测；牛奶和鱼塘水可以直接进行检测；乳粉5倍加入PBS溶解；蜂蜜5倍稀释。该免疫亲和凝胶柱检测食品样品不需要任何有机溶剂和复杂的前处理,及大型仪器的辅助,添加3个不同浓度的伏马毒素(氯霉素)标品,用ELISA方法进行了验证,检测结果一致,表明试验所建立的方法具有很好的准确性,满足快速检测的要求。
[Abstract]:The detection of FB1 and CAP residues in food was systematically studied by an immuno-affinity gel column. The agarose gel coupled with fumaroxin antibody or chloramphenicol antibody was used to prepare its respective antibody glue. Horseradish catalase (HRP) antibody was coupled to prepare the HRP gel, and the agarose gel activated by hydrogen bromide was blocked to prepare the sealant. The sealant and antibody gel were mixed to prepare the detection layer, and the HRP gel was mixed to form the contrast layer. The test column was mounted. The factors affecting the experiment (ionic strength, pH value, etc.) were optimized, and a time-saving and high sensitivity was established. Visual detection method can be used to detect fumaroxin and chloramphenicol in food qualitatively and quantitatively. For Fumatoxin B1 immuno-affinity gel column: the mixture of sealant and antibody glue at 1:10 can achieve good visual color results, and the control layer was prepared by mixing sealant and antibody glue 1:90. The FB1-HRP was diluted with 1: 8000 to detect the gel column. The minimum detection limit (LOD) of the FB1-HRP gel column was 40 渭 g 路L ~ (-1) 路L ~ (-1). The residue of fumarin in beer, liquor, cornmeal and cereal milk was determined: beer and liquor were diluted 5 times directly, corn flour was diluted by 5 times after PBS extraction, and the upper layer solution of cereal milk was diluted by 10 times. For chloramphenicol immunoaffinity gel detection column: the mixture of sealant and antibody glue at 1:20 can achieve good visual color result, and the control layer was prepared by mixing the sealant and antibody glue 1: 100. The CAP-HRP was diluted at 1: 200 000 to detect the gel column. The LOD of the established gel column was 1 渭 g / L. The residues of chloramphenicol in milk, milk powder, fish pond water and honey were detected; milk and fish pond water could be directly detected; milk powder was dissolved by PBS; honey was diluted by 5 times. The immunoaffinity gel column was used to detect food samples without any organic solvent, complex pretreatment, and the aid of large scale instrument. Three different concentrations of chloramphenicol (chloramphenicol) were added to food samples, and the results were verified by Elisa. The results show that the proposed method has good accuracy and meets the requirement of rapid detection.
【学位授予单位】：天津科技大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R155.5

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