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基于三重标记多重扩增的食源性细菌同步电化学生物磁传感

Simultaneous electrochemical magneto genosensing of foodborne bacteria based on triple-tagging multiplex amplification.

机构信息

Grup de Sensors i Biosensors, Departament de Química, Universitat Autònoma de Barcelona, 08193 Cerdanyola del Vallès (Bellaterra), Spain.

Applied Enzyme Technology Ltd., Gwent Group Ltd., Monmouth House, Mamhilad Park, Pontypool NP4 OHZ, UK.

出版信息

Biosens Bioelectron. 2015 Dec 15;74:652-9. doi: 10.1016/j.bios.2015.07.008. Epub 2015 Jul 9.

Abstract

Simultaneous detection of Salmonella enterica, Listeria monocytogenes and Escherichia coli based on triple-tagging multiplex PCR and electrochemical magneto genosensing on silica magnetic particles is reported. A set of tagging primers were selected for the specific amplification of yfiR (375 bp), hlyA (234 bp) and eaeA (151bp), being one of the primers for each set labelled with fluorescein, biotin and digoxigenin coding for S. enterica, L. monocytogenes and E. coli, respectively. Afterwards, electrochemical magneto genosensing of the bacteria was achieved by using silica magnetic particles as a carrier and three different electrochemical reporters, specific for each pathogen. This method was able to clearly distinguish among the pathogenic bacteria tested within 50 min, with detection limits ranging from 12 to 46 pg μL(-1).

摘要

基于三重标记多重 PCR 和基于硅基磁性粒子的电化学生磁传感同时检测沙门氏菌、李斯特菌和大肠杆菌。本研究选择了一组标记引物,用于特异性扩增 yfiR(375bp)、hlyA(234bp)和 eaeA(151bp),其中每个引物的一条链标记有荧光素、生物素和地高辛,分别用于编码沙门氏菌、李斯特菌和大肠杆菌。随后,通过使用硅基磁性粒子作为载体和三种针对每种病原体的不同电化学生物传感器,实现了对细菌的电化学磁传感。该方法能够在 50 分钟内清晰地区分所测试的病原菌,检测限范围为 12 至 46pgμL(-1)。

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