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用于黄曲霉毒素B1放大光学检测的免疫金可控生长

Controlled growth of immunogold for amplified optical detection of aflatoxin B1.

作者信息

Wang Xu, Niessner Reinhard, Knopp Dietmar

机构信息

Institute of Hydrochemistry, Chair for Analytical Chemistry, Technische Universität München, Marchioninistrasse 17, D-81377 München, Germany.

出版信息

Analyst. 2015 Mar 7;140(5):1453-8. doi: 10.1039/c4an02281e. Epub 2015 Jan 20.

Abstract

A simple, sensitive and cost-effective method for the analysis of the mycotoxin aflatoxin B1 (AFB1) has been established based on controlled growth of immunogold. AFB1-BSA conjugate modified magnetic beads were employed as capture probe and anti-AFB1 antibody-coated gold colloids were used as detection probe for the immunological recognition of AFB1, as well as for signal transduction. The immune recognition event is converted into the gold enlargement signal which can be quantitatively measured by UV-vis spectroscopy. The autocatalytic enlargement of immunogold was conducted in aqueous solution containing chloroauric acid, hexadecyltrimethylammonium bromide and ascorbic acid. The reaction could be stopped by the addition of sodium thiosulfate. The final absorbance and resonance light scattering intensity were highly dependent on immunogold concentration. After gold enhancement, the sensitivity of the immunoassay was improved and total assay time reduced to 1 h. Under optimized conditions, the linear range and lower detection limit was 0.01-1 ng mL(-1) and 7 pg mL(-1), respectively. The proposed method offers great promise for sensitive detection of other mycotoxins and organic pollutants.

摘要

基于免疫金的可控生长,建立了一种简单、灵敏且经济高效的黄曲霉毒素B1(AFB1)分析方法。采用AFB1 - BSA偶联物修饰的磁珠作为捕获探针,抗AFB1抗体包被的金胶体作为检测探针,用于AFB1的免疫识别及信号转导。免疫识别事件转化为金放大信号,可通过紫外 - 可见光谱进行定量测定。免疫金的自催化放大在含有氯金酸、十六烷基三甲基溴化铵和抗坏血酸的水溶液中进行。反应可通过加入硫代硫酸钠终止。最终吸光度和共振光散射强度高度依赖于免疫金浓度。金增强后,免疫分析的灵敏度提高,总分析时间缩短至1小时。在优化条件下,线性范围和最低检测限分别为0.01 - 1 ng mL(-1)和7 pg mL(-1)。该方法为其他霉菌毒素和有机污染物的灵敏检测提供了广阔前景。

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