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基于 O-羧甲基壳聚糖表面修饰的 Fe3O4 纳米粒子的电化学阻抗免疫传感器用于腹泻患者粪便中弯曲杆菌属空肠的无标记检测

An electrochemical impedimetric immunosensor for label-free detection of Campylobacter jejuni in diarrhea patients' stool based on O-carboxymethylchitosan surface modified Fe3O4 nanoparticles.

机构信息

College of Bioscience and Biotechnology, Yangzhou University, Yangzhou, PR China.

出版信息

Biosens Bioelectron. 2010 Jan 15;25(5):1204-11. doi: 10.1016/j.bios.2009.10.036. Epub 2009 Nov 4.

Abstract

A novel electrochemical impedimetric immunosensor based on O-carboxymethylchitosan surface modified Fe(3)O(4) nanoparticles (denoted as OCMCS-Fe(3)O(4) nanoparticles) was developed for rapid detection of Campylobacter jejuni, which is becoming the most common cause of gastroenteritis in developed countries and raising major public health concerns worldwide. In the present study, anti-FlaA monoclonal antibodies 2D12 (denoted as 2D12McAbs) were immobilized on OCMCS-Fe(3)O(4) nanoparticles. The detection was performed by measuring relative change in impedance before and after 2D12McAbs-Campylobacter jejuni reaction with the technique of electrochemical impedance spectroscopy. Under the optimized conditions, the relative change in impedance was proportional to the logarithmic value of Campylobacter jejuni concentrations in the range of 1.0x10(3) to 1.0x10(7) CFU/mL (r=0.991). The advantages of the OCMCS-Fe(3)O(4) nanoparticle-based immunosensor are simplicity of use, fast response, wide linear range, acceptable reproducibility and long stability. Moreover, the immunosensor could be regenerated by being treated with glycine-HCl buffer solution (pH 2.8). We demonstrate the convenient application of the novel immunosensor for the detection of Campylobacter jejuni in diarrhea patients' stool samples.

摘要

一种基于 O-羧甲基壳聚糖表面修饰的四氧化三铁纳米粒子(记为 OCMCS-Fe3O4 纳米粒子)的新型电化学阻抗免疫传感器,用于快速检测空肠弯曲菌,空肠弯曲菌是发达国家最常见的胃肠炎病原体,引起了全球主要的公共卫生关注。在本研究中,将抗 FlaA 单克隆抗体 2D12(记为 2D12McAbs)固定在 OCMCS-Fe3O4 纳米粒子上。通过电化学阻抗谱技术测量 2D12McAbs-空肠弯曲菌反应前后的阻抗相对变化来进行检测。在优化条件下,阻抗的相对变化与空肠弯曲菌浓度的对数在 1.0x10(3)到 1.0x10(7)CFU/mL 的范围内成正比(r=0.991)。基于 OCMCS-Fe3O4 纳米粒子的免疫传感器具有使用简便、响应快速、线性范围宽、重现性好、稳定性长等优点。此外,该免疫传感器可以用甘氨酸-HCl 缓冲溶液(pH 2.8)处理进行再生。我们证明了这种新型免疫传感器在检测腹泻患者粪便样本中空肠弯曲菌方面的便捷应用。

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