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高灵敏度和选择性电化学生物传感器:使用亲和肽作为分子结合物对人诺如病毒进行无标记检测。

High sensitive and selective electrochemical biosensor: Label-free detection of human norovirus using affinity peptide as molecular binder.

机构信息

Department of Pharmaceutical Engineering, Daegu Haany University, Gyeongsan 38610, Republic of Korea.

Department of Chemistry, Chung-Ang University, Seoul 06974, Republic of Korea.

出版信息

Biosens Bioelectron. 2017 Jan 15;87:164-170. doi: 10.1016/j.bios.2016.08.031. Epub 2016 Aug 12.

Abstract

Norovirus is known as the major cause of highly infection for gastrointestinal tracts. In this study, robust and highly sensitive biosensors for detecting human norovirus by employing a recognition affinity peptide-based electrochemical platform were described. A series of amino acid-substituted and cysteine-incorporated recognition peptides isolated from evolutionary phage display technique was chemically synthesized and immobilized to a gold sensor layer, the detection performance of the gold-immobilized synthetic peptide-based sensor system was assessed using QCM, CV and EIS. Using EIS, the limit of detection with Noro-1 as a molecular binder was found to be 99.8nM for recombinant noroviral capsid proteins (rP2) and 7.8copies/mL for human norovirus, thereby demonstrating a high degree of sensitivity for their corresponding targets. These results suggest that a biosensor which consists of affinity peptides as a molecular binder and miniaturized microdevices as diagnostic tool could be served as a new type of biosensing platform for point-of-care testing.

摘要

诺如病毒是一种主要的胃肠道高度传染性病原体。本研究采用基于识别亲和肽的电化学生物传感器,描述了用于检测人类诺如病毒的强大且高灵敏度的生物传感器。从进化噬菌体展示技术中分离出的一系列氨基酸取代和半胱氨酸掺入的识别肽被化学合成并固定在金传感器层上,使用 QCM、CV 和 EIS 评估金固定合成肽基传感器系统的检测性能。使用 EIS,发现作为分子结合物的诺如 1 对重组诺如病毒衣壳蛋白 (rP2) 的检测限为 99.8nM,对人类诺如病毒的检测限为 7.8 拷贝/mL,从而对其相应的靶标表现出高度的灵敏度。这些结果表明,由亲和肽作为分子结合物和微型化微器件作为诊断工具组成的生物传感器可以作为一种新型的即时检测生物传感平台。

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