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基于芯片的电化学生物传感器用于灵敏、无标记检测隐孢子虫。

On-chip-based electrochemical biosensor for the sensitive and label-free detection of Cryptosporidium.

机构信息

School of Engineering, University of British Columbia, 333 University Way, Kelowna, BC, V1V1V7, Canada.

出版信息

Sci Rep. 2022 Apr 28;12(1):6957. doi: 10.1038/s41598-022-10765-0.

Abstract

Cryptosporidium, an intestinal protozoan pathogen, is one of the leading causes of death in children and diarrhea in healthy adults. Detection of Cryptosporidium has become a high priority to prevent potential outbreaks. In this paper, a simple, easy to fabricate, and cost-effective on-chip-based electrochemical biosensor has been developed for the sensitive and label-free detection of Cryptosporidium oocysts in water samples. The sensor was fabricated using standard lithography using a mask with a 3-electrode design and modified by self-assembling a hybrid of a thiolated protein/G and the specific anti-Cryptosporidium monoclonal antibodies (IgG3). The electrochemical impedance spectroscopy (EIS) was employed to quantitate C. parvum in the range of 0 to 300 oocysts, with a detection limit of approximately 20 oocysts/5 µL. The high sensitivity and specificity of the developed label-free electrochemical biosensor suggest that this novel platform is a significant step towards the development of fast, real-time, inexpensive and label-free sensing tool for early warning and immediate on-site detection of C. parvum oocysts in water samples, as compared to the traditional methods (such as PCR and microscopy). Furthermore, under optimized conditions, this label-free biosensor can be extended to detect other analytes and biomarkers for environmental and biomedical analyses.

摘要

隐孢子虫是一种肠道原生动物病原体,是导致儿童死亡和健康成年人腹泻的主要原因之一。检测隐孢子虫已成为预防潜在疫情的首要任务。在本文中,我们开发了一种简单、易于制造且具有成本效益的基于芯片的电化学生物传感器,用于灵敏且无标记地检测水样中的隐孢子虫卵囊。该传感器采用标准光刻技术,使用带有 3 电极设计的掩模制造,并通过自组装硫醇化蛋白/G 和特异性抗隐孢子虫单克隆抗体(IgG3)的混合物进行修饰。电化学阻抗谱(EIS)用于定量检测 0 至 300 个隐孢子虫卵囊,检测限约为 20 个卵囊/5μL。所开发的无标记电化学生物传感器具有高灵敏度和特异性,表明该新型平台是朝着开发快速、实时、廉价和无标记传感工具迈出的重要一步,可用于早期预警和即时现场检测水样中的隐孢子虫卵囊,与传统方法(如 PCR 和显微镜)相比。此外,在优化条件下,这种无标记生物传感器可以扩展到检测其他用于环境和生物医学分析的分析物和生物标志物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6fba/9051104/abc07b1fd73e/41598_2022_10765_Fig1_HTML.jpg

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