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

中文关键词:  卡那霉素  戊二醛  PAMAM  表面修饰

英文关键词:immunoassay method  milk  kanamycin  glutaraldehyde  polyamidoamine  surface modified

中文摘要:

      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。

英文摘要:

      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.