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基于寡核苷酸的流通式电化学生物传感器检测 。

Flow-Through Electrochemical Biosensor for the Detection of Using Oligonucleotides.

机构信息

United States Department of Agriculture, Agriculture Research Service, Eastern Regional Research Center, 600 East Mermaid Lane, Wyndmoor, PA 19038, USA.

出版信息

Sensors (Basel). 2021 May 28;21(11):3754. doi: 10.3390/s21113754.

Abstract

Consumption of food contaminated by can result in Listeriosis, an illness with hospitalization rates of 94% and mortality rates up to 30%. As a result, U.S. regulatory agencies governing food safety retain zero-tolerance policies for . However, detection at such low concentrations often requires strategies such as increasing sample size or culture enrichment. A novel flow-through immunoelectrochemical biosensor has been developed for O157:H7 detection in 1 L volumes without enrichment. The current work further augments this biosensor's capabilities to (1) include detection of and (2) accommodate genetic detection to help overcome limitations based upon antibody availability and address specificity errors in phenotypic assays. Herein, the conjugation scheme for oligo attachment and the conditions necessary for genetic detection are laid forth while results of the present study demonstrate the sensor's ability to distinguish DNA from with a limit of detection of ~2 × 10 cells/mL, which agrees with prior studies. Total time for this assay can be constrained to <2.5 h because a timely culture enrichment period is not necessary. Furthermore, the electrochemical detection assay can be performed with hand-held electronics, allowing this platform to be adopted for near-line monitoring systems.

摘要

食用受污染的食物可能导致李斯特菌病,这种疾病的住院率为 94%,死亡率高达 30%。因此,负责食品安全的美国监管机构对李斯特菌保持零容忍政策。然而,在如此低的浓度下进行检测通常需要增加样本量或培养富集等策略。已经开发出一种新型的流通免疫电化学生物传感器,用于在不进行富集的情况下检测 1 L 体积中的 O157:H7。目前的工作进一步增强了该生物传感器的功能,(1)包括检测和(2)适应基因检测,以帮助克服基于抗体可用性的限制,并解决表型测定中的特异性错误。本文阐述了寡核苷酸连接的缀合方案以及进行基因检测所需的条件,而本研究的结果表明,该传感器能够区分 和 ,检测限约为~2×10 个细胞/mL,与先前的研究一致。由于不需要及时进行培养富集,因此整个检测时间可以限制在<2.5 小时内。此外,电化学检测可以使用手持式电子设备进行,从而使该平台能够被用于在线监测系统。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/796e/8198859/519981df4023/sensors-21-03754-g001.jpg

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