摘要
试验旨在建立灵敏、快速、特异的氯霉素(CAP)检测方法,基于免疫亲和色谱技术和酶联免疫技术,研究了一种新型快速检测虾肉中CAP的免疫亲和色谱柱(ICTC)检测方法。采用溶胶-凝胶(sol-gel)法制备ICTC柱的检测层和控制层,优化检测层、控制层、酶标物(CAP-HRP)稀释度等条件,制备ICTC柱。测定ICTC柱的灵敏度、特异性等性能参数,并对虾肉样本前处理方法和基质干扰效应进行优化。本研究确定ICTC柱条件为:检测层,anti-CAP sol-gel∶空白sol-gel为10∶1 000(V/V);控制层,anti-HRP sol-gel∶空白sol-gel为13∶1 000(V/V);CAP-HRP的稀释度为1∶105。采用检测层消色法判定结果,检测时间10min,PBS缓冲液中CAP的检测限为0.5μg/L,该方法与甲砜霉素、氟苯尼考、氟苯尼考胺同类药物或代谢产物均无可见交叉反应,在虾肉样本中的检测限为0.75μg/kg。本研究建立的ICTC柱检测方法快速、灵敏,使用方便,为现场快速检测虾肉中CAP残留提供了新的技术手段。
The aim of this study was to establish a sensitive,rapid and specific method for the detection of chloramphenicol(CAP).Based on immunoaffinity chromatography and enzyme-linked immunosorbent assay,a novel immunoaffinity chromatography test column(ICTC)for the rapid detection of CAP in shrimp was developed.The detection layer and control layer of ICTC column were prepared by sol-gel method,and the conditions of detection layer,control layer and CAP-HRP dilution factor were optimized.The performance parameters such as sensitivity and specificity of the ICTC column were determined,and the pretreatment method and matrix interference effect of the shrimp sample were optimized.This study determined that the ICTC column conditions were as follow:The detection layer,anti-CAP sol-gel∶blank sol-gel was 10∶1 000(V/V);The control layer,anti-HRP sol-gel∶blank sol-gel was 13∶1 000(V/V);CAP-HRP dilution was 1∶10 5.The detection result was determined by the detection layer decolorization method.The detection time was 10 min,and the detection limit of CAP in PBS buffer was 0.5μg/L.The method had no visible cross-reaction with thiabendazole,florfenicol,florfenicol similar drugs or metabolites.The detection limit of this method in shrimp samples was 0.75μg/kg.The ICTC column detection method established in this study was fast,sensitive and easy to use,and provided a new technical methods for rapid detection of CAP residues in shrimp on site.
作者
吴穗勤
黄珊
高铖
孟辉
WU Suiqin;HUANG Shan;GAO Cheng;MENG Hui(School of Environment and Safety Engineering,Jiangsu University, Zhenjiang 212013,China)
出处
《中国畜牧兽医》
CAS
北大核心
2018年第10期2894-2901,共8页
China Animal Husbandry & Veterinary Medicine
基金
国家自然科学基金(31502114)
江苏省大学生创新项目(201610299018Z)
作者简介
吴穗勤(1994-),女,广东汕头人,学士,研究方向:免疫分析方法,E-mail:1421214118@qq.com;通信作者:孟辉(1985-),男,山东兖州人,博士,研究方向:免疫分析方法,E-mail:menghui@ujs.edu.cn
