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基于特异性适体的牛奶样品中黄曲霉毒素 M1 检测的微流控装置的制作。

Fabrication of microfluidic device for Aflatoxin M1 detection in milk samples with specific aptamers.

机构信息

DBT- National Institute of Animal Biotechnology, Hyderabad, 500032, India.

Department of Biotechnology, JNTUA College of Engineering, Andhra Pradesh, 516390, India.

出版信息

Sci Rep. 2020 Mar 13;10(1):4627. doi: 10.1038/s41598-020-60926-2.

Abstract

This study describes the colorimetric detection of aflatoxin M1 (Afl M1) in milk samples using a microfluidic paper-based analytical device (µPAD). Fabrication of µPADs was done using a simple and quick approach. Each μPAD contained a detection zone and a sample zone interconnected by microchannels. The colorimetric assay was developed using unmodified AuNPs as a probe and 21-mer aptamer as a recognition molecule. The free aptamers were adsorbed onto the surface of AuNPs in absence of Afl M1, even at high salt concentrations. The salt induced aggregation of specific aptamers occurred in presence of Afl M1. Under optimum conditions, the analytical linear range was found to be 1 µM to 1 pM with limit of detection 3 pM and 10 nM in standard buffer and spiked milk samples respectively. The proposed aptamer based colorimetric assay was repeatable, quick, selective, and can be used for on-site detection of other toxins in milk and meat samples.

摘要

本研究描述了使用微流控纸基分析装置(µPAD)对牛奶样品中的黄曲霉毒素 M1(Afl M1)进行比色检测。µPAD 的制造采用了简单快捷的方法。每个 μPAD 包含一个检测区和一个样品区,通过微通道相互连接。比色测定法使用未经修饰的 AuNPs 作为探针和 21 个碱基的适体作为识别分子。在不存在 Afl M1 的情况下,游离适体被吸附在 AuNPs 的表面上,即使在高盐浓度下也是如此。在存在 Afl M1 的情况下,特定适体的盐诱导聚集发生。在最佳条件下,分析线性范围在 1 µM 至 1 pM 之间,检测限分别为 3 pM 和 10 nM,在标准缓冲液和加标牛奶样品中。基于适体的比色测定法具有可重复性、快速、选择性,可用于现场检测牛奶和肉类样品中的其他毒素。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bdef/7070014/67d697383bbb/41598_2020_60926_Sch1_HTML.jpg

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