基于表面修饰的高灵敏度免疫分析方法检测牛奶中的卡那霉素  

Highly sensitivity immunoassay based on surface modification for the detection of kanamycin in milk

作　　者：顾鑫凯 朱宝成 王亚妮 黄睿 曾昆 张祯 GU Xin-Kai;ZHU Bao-Cheng;WANG Ya-Ni;HUANG Rui;ZENG Kun;ZHANG Zhen(School of Environmental and Safety Engineering,Jiangsu University,Zhenjiang 212001,China)

机构地区：[1]江苏大学环境与安全工程学院,镇江212001

出　　处：《食品安全质量检测学报》2024年第4期59-67,共9页Journal of Food Safety and Quality

基　　金：国家自然科学基金项目(22176075);江苏省农业科技自主创新资金项目(CX(21)3173)。

摘　　要：目的为了提高卡那霉素的检测灵敏度,将卡那霉素分子直接共价固化在酶标板上,由此开发一种直接免疫分析方法检测卡那霉素。方法本研究采用戊二醛以及聚乙二胺(polyamidoamine,PAMAM)-戊二醛两种方式,对酶标板进行表面修饰,同时通过酶联免疫吸附实验(enzyme-linked immunosorbent assays,ELISA)比较两种方式的差异,并对牛奶样本中卡那霉素进行检测。结果基于戊二醛表面修饰的ELISA检出限为0.219 ng/mL,线性范围为2.251~87.791 ng/mL;基于PAMAM-戊二醛表面修饰的ELISA检出限为0.119 ng/mL,线性范围为1.348~20.414 ng/mL。此两种方法比常规ELISA方法灵敏度分别提高5倍和10倍。将基于PAMAM-戊二醛表面修饰的ELISA应用于牛奶样本的检测,发现卡那霉素的检出率为12.5%,最高检出质量浓度为1.12 ng/mL。结论本研究采用的戊二醛以及PAMAM-戊二醛两种方式修饰酶标板后大大提高了免疫检测的灵敏度,为食品中小分子污染物的快速筛查提供了新的思路。Objective To improve kanamycin detection sensitivity,develope a direct immunoassay approach by directly covalently anchoring kanamycin molecules to the surface of a modified plate.Methods The surface of the enzyme-labeled plates was altered in this investigation using glutaraldehyde and polyamidoamine(PAMAM)-glutaraldehyde.Meanwhile,enzyme-linked immunosorbent assays(ELISA)were utilized to compare the two approaches.Kanamycin has been identified in milk samples.Results For glutaraldehyde surface modified ELISA,the linear range was 2.251-87.791 ng/mL and the limit of detection was 0.219 ng/mL.For PAMAM-glutaraldehyde surface modified ELISA,the linear range was 1.348-20.414 ng/mL and the limit of detection was 0.119 ng/mL.The sensitivities of the two methods improved 5-fold and 10-fold compared with conventional ELISA.Using an ELISA based on PAMAM-glutaraldehyde to analyze milk samples,the detection rate of kanamycin was 12.5%and the highest detection mass concentration was 1.12 ng/mL.Conclusion Glutaraldehyde and PAMAM-glutaraldehyde modified enzyme plate in this study greatly improve the sensitivity of immune detection,and provide a novel concept for the quick screening of contaminants in food.

关 键 词：免疫分析方法 牛奶 卡那霉素 戊二醛 聚乙二胺 表面修饰 

分 类 号：TS252.7[轻工技术与工程—农产品加工及贮藏工程]