[关键词]
[摘要]
本文提出了一种基于抗体功能化纳米氧化铜信号示踪的比色免疫分析检测黄曲霉毒素B1。本方法的建立是基于羧基化磁珠表面共价结合抗体与黄曲霉毒素B1高效结合后,再与静电吸附在纳米氧化铜表面的抗体特异性识别,定量结合的氧化铜标记物可经酸溶解释放出高浓度的铜离子,加入腙类铜离子显色剂即可生成红色络合物,紫外-可见吸收光谱法测定该红色溶液的吸光度,或直接通过目视比色法即可方便地实现免疫分析信号转导从而实现快速检测黄曲霉毒素B1。结果表明,在培育时间为15 min及加入显色剂浓度为10 μmol/L时,该方法具有较高的检测灵敏度并在0.01~100 ng/mL范围内具有良好的相关性,检测下限是0.035 ng/mL。此外,该方法已经成功应用于分析大米样品中的黄曲霉毒素B1,其结果与GB 5009.22中的方法相一致。
[Key word]
[Abstract]
A method of colorimetric immunoassay based on functionalization of copper oxide nanoparticles was established for the rapid and sensitive detection of Alflatoxin B1 (AFB1), a tumor biomarker. The method was established based on that the carboxyl covalent bonding magnetic bead surface antibody was highly bound to aflatoxin B1, and then was specifically recognized by the electrostatic adsorption on the surface of the nano copper oxide, and the quantitative bound copper oxide marker could release high concentration of copper ions by acid solution, and then the copper ion chromogenic agent of the hydrazone could generate red complexes. The absorbance of the red solution was determined by the ultraviolet visible absorption spectrometry, or directly through the visual colorimetric method, which could easily realize the signal transduction of immune analysis so as to realize rapid detection of aflatoxin B1. The results showed that the method had a high detection sensitivity and a good correlation in the range of 0.01 to 100 ng/mL when the incubation time was 15 min and the oncentration of chromogenic agent was 10 μmol/L. In addition, this method has been successfully applied to the analysis of alflatoxin B1 in rice samples, and the results were consistent with those in GB 5009.22.
[中图分类号]
[基金项目]
国家自然科学基金项目(No.21475033)