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使用树枝状聚合物作为多把手模板对微流控装置进行适体表面功能化及其在食源性病原体敏感检测中的应用。

Aptamer surface functionalization of microfluidic devices using dendrimers as multi-handled templates and its application in sensitive detections of foodborne pathogenic bacteria.

机构信息

Department of Chemical and Biological Engineering, University of Ottawa, Ottawa, Ontario, K1N 6N5, Canada.

Nanchang Institute of Technology, 901 Yingxiong Road, Nanchang, Jiangxi, 330044, China.

出版信息

Anal Chim Acta. 2019 May 16;1056:96-107. doi: 10.1016/j.aca.2019.01.035. Epub 2019 Jan 29.

DOI:10.1016/j.aca.2019.01.035
PMID:30797466
Abstract

A microfluidic system that incorporates both dendrimers and aptamers to detect E. coli O157:H7 is developed. To achieve this, generation 7-polyamidoamine dendrimers were immobilized onto the detection surfaces of PDMS microfluidic channels; subsequently aptamers against E. coli O157:H7 were conjugated onto the microchannel surfaces via the immobilized dendrimers as templates. Surface modifications were characterized by FTIR, XPS, water contact angles, fluorescence microscopy and AFM to confirm the success of each surface modification steps. The efficacy of this simple microchannel in detection was investigated using E. coli O157:H7 spiked samples. Our results showed that this interesting approach significantly increased the amount of aptamers available on the microfluidic channel surfaces to capture E. coli O157:H7 cells to allow sensitive detection, which in turn resulted in detections of E. coli O157:H7 cells at a low limit of detection of 10 cells mL. The results also demonstrated that in comparison with the generation 4-polyamidoamine dendrimers (G4) modified microchannels, those modified with G7 showed enhanced detection signals, improved target capturing efficiencies, and at higher throughput. This simple whole cell detection design has not been reported in the literature and it is an interesting and effective approach to developing a sensitive and rapid detection platform for foodborne pathogenic bacteria.

摘要

一种结合树状高分子和适体来检测大肠杆菌 O157:H7 的微流控系统被开发出来。为了实现这一目标,第 7 代聚酰胺-胺树状高分子被固定在 PDMS 微流控通道的检测表面上;随后,针对大肠杆菌 O157:H7 的适体通过固定在微通道表面上的树状高分子作为模板被连接到微通道表面上。表面修饰通过傅里叶变换红外光谱(FTIR)、X 射线光电子能谱(XPS)、水接触角、荧光显微镜和原子力显微镜(AFM)进行表征,以确认每个表面修饰步骤的成功。通过使用大肠杆菌 O157:H7 污染的样品来研究这个简单微通道在检测中的效果。我们的结果表明,这种有趣的方法显著增加了微流控通道表面上可用的适体数量,以捕获大肠杆菌 O157:H7 细胞,从而实现敏感检测,这反过来又导致可以在低检测限 10 个细胞 mL 下检测到大肠杆菌 O157:H7 细胞。结果还表明,与第 4 代聚酰胺-胺树状高分子(G4)修饰的微通道相比,用 G7 修饰的微通道显示出增强的检测信号、提高的目标捕获效率和更高的通量。这种简单的全细胞检测设计在文献中尚未报道,它是开发用于食源性病原体的敏感和快速检测平台的一种有趣且有效的方法。

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