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基于石英晶体微天平的生物催化沉积扩增法检测黄曲霉毒素B1

Biocatalyzed deposition amplification for detection of aflatoxin B1 based on quartz crystal microbalance.

作者信息

Jin Xuefang, Jin Xiaoyong, Liu Xueping, Chen Liguo, Jiang Jianhui, Shen Guoli, Yu Ruqin

机构信息

State Key Laboratory of Chemo/Biosensing and Chemometrics, Chemistry and Chemical Engineering College, Hunan University, Changsha 410082, PR China.

出版信息

Anal Chim Acta. 2009 Jul 10;645(1-2):92-7. doi: 10.1016/j.aca.2009.04.041. Epub 2009 May 5.

Abstract

An ultrasensitive piezoelectric method for the detection of the aflatoxin B1 (AFB1) based on the indirect competitive immunoassay and the biocatalyzed deposition amplification has been developed. In this method, the quartz crystal surface was coated with a self-assembled monolayer of 3-mercaptopropionic acid (MPA) for covalently immobilization of the BSA-AFB1 conjugate, which could compete with the free AFB1 for binding to the anti-AFB1 antibody (MsIgG). After the competitive immunoreaction, the horseradish peroxidase (HRP) labeled goat anti-mouse IgG (G-Anti-MsIgG) was introduced into the detection cell to combine with the anti-AFB1 antibody on the crystal surface. The enzyme labeled G-Anti-MsIgG as a biocatalyst could accelerate the oxidation of 4-chloro-1-naphthol by H2O2 to yield the insoluble product benzo-4-chlorohexadienone on the surface of quartz crystal microbalance (QCM), resulting in a mass increase that was reflected by a decrease in the resonance frequency of the QCM. The proposed approach could allow for the determination of AFB(1) in the concentration range of 0.01-10.0 ng mL(-1). Furthermore, several artificially contaminated milk samples were analyzed with good recoveries obtained, which demonstrated the suitability of the proposed method for detecting AFB1.

摘要

基于间接竞争免疫分析和生物催化沉积放大技术,开发了一种用于检测黄曲霉毒素B1(AFB1)的超灵敏压电方法。在该方法中,石英晶体表面涂覆有3-巯基丙酸(MPA)自组装单分子层,用于共价固定牛血清白蛋白-AFB1偶联物,该偶联物可与游离AFB1竞争结合抗AFB1抗体(MsIgG)。竞争免疫反应后,将辣根过氧化物酶(HRP)标记的山羊抗小鼠IgG(G-Anti-MsIgG)引入检测池中,与晶体表面的抗AFB1抗体结合。酶标记的G-Anti-MsIgG作为生物催化剂,可加速H2O2氧化4-氯-1-萘酚,在石英晶体微天平(QCM)表面产生不溶性产物苯并-4-氯己二烯酮,导致质量增加,这通过QCM共振频率的降低来反映。所提出的方法能够测定浓度范围为0.01-10.0 ng mL(-1)的AFB(1)。此外,对几个人工污染的牛奶样品进行了分析,回收率良好,这证明了所提出的方法适用于检测AFB1。

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