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基于纳米酶和适体的黄曲霉毒素 B1 免疫吸附测定法。

Nanozyme and aptamer- based immunosorbent assay for aflatoxin B1.

机构信息

National "111" Center for Cellular Regulation and Molecular Pharmaceutics, Key Laboratory of Fermentation Engineering (Ministry of Education), College of Bioengineering and Food, Hubei University of Technology, Wuhan, Hubei 430068, PR China; College of Food Science and Technology, Huazhong Agricultural University, Wuhan, Hubei 430070, PR China.

National "111" Center for Cellular Regulation and Molecular Pharmaceutics, Key Laboratory of Fermentation Engineering (Ministry of Education), College of Bioengineering and Food, Hubei University of Technology, Wuhan, Hubei 430068, PR China; College of Food Science and Technology, Huazhong Agricultural University, Wuhan, Hubei 430070, PR China.

出版信息

J Hazard Mater. 2020 Nov 15;399:123154. doi: 10.1016/j.jhazmat.2020.123154. Epub 2020 Jun 11.

Abstract

Traditional enzyme-linked immunosorbent assay (ELISA) suffers from the limitations of relatively low sensitivity and stability, and enzyme-labelled antibodies are hard to be prepared and purified. Based on a nanozyme, an aptamer and FeO magnetic nanoparticles (MNP), a nanozyme and aptamer-based immunosorbent assay (NAISA) was developed for aflatoxin B1 (AFB1) detection with simpler operation and separation. In this work, mesoporous SiO/Au-Pt (m-SAP) were prepared to act as signal labels, which showed high catalase-like activity and was denoted as nanozyme. Aptamer was adopted to specifically recognize with AFB1, and MNP facilitated to realize magnetic separation. To verify the performance of NAISA, traditional ELISA (t-ELISA) and enhanced ELISA (e-ELISA) using MNP and m-SAP nanozyme were applied in AFB1 detection. The NAISA method showed the lowest limit of detection (LOD) with 5 pg mL (n = 3, ±4.2 %), 600 and 12-fold lower than that of t-ELISA (3 ng mL) and e-ELISA (0.06 ng mL), respectively. In the interference tests, AFB1 can be identified among six different interfering substances. The NAISA method, thus, can be of great importance as it allows selective and sensitive AFB1 detection, while providing the simplicity of use and need for screening hazardous materials.

摘要

传统的酶联免疫吸附测定(ELISA)存在灵敏度和稳定性相对较低的局限性,并且酶标记抗体难以制备和纯化。基于纳米酶、适体和 FeO 磁性纳米粒子(MNP),开发了一种基于纳米酶和适体的免疫吸附测定法(NAISA),用于检测黄曲霉毒素 B1(AFB1),具有更简单的操作和分离。在这项工作中,制备了介孔 SiO/Au-Pt(m-SAP)作为信号标签,其具有高过氧化物酶样活性,被称为纳米酶。适体被用来特异性识别 AFB1,而 MNP 则有助于实现磁性分离。为了验证 NAISA 的性能,采用传统 ELISA(t-ELISA)和使用 MNP 和 m-SAP 纳米酶的增强 ELISA(e-ELISA)来检测 AFB1。NAISA 法的检测限(LOD)最低,为 5 pg mL(n = 3,±4.2%),比 t-ELISA(3 ng mL)和 e-ELISA(0.06 ng mL)分别低 600 倍和 12 倍。在干扰试验中,AFB1 可以在六种不同的干扰物质中被识别。因此,NAISA 法可以作为一种重要的方法,因为它允许选择性和灵敏性地检测 AFB1,同时提供简单的使用和筛选有害物质的需要。

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