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一种基于冰晶状金纳米结构的新型超灵敏电化学 DNA 生物传感器,用于检测粪肠球菌基因序列。

A novel and ultrasensitive electrochemical DNA biosensor based on an ice crystals-like gold nanostructure for the detection of Enterococcus faecalis gene sequence.

机构信息

Nanomedicine and Nanobiology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.

Nanomedicine and Nanobiology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran; Department of Medical Physics, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.

出版信息

Colloids Surf B Biointerfaces. 2018 Jun 1;166:245-253. doi: 10.1016/j.colsurfb.2018.03.025. Epub 2018 Mar 20.

Abstract

Bacteria, parasites and viruses are found widely in the environment as potential pathogens, and can be the source of infections. Therefore, sensitive and rapid methods for identification of the pathogens are required to achieve a better quality of life. Enterococcus faecalis commonly colonizes and threatens human health. In the present study, we demonstrate the fabrication of a novel electrochemical DNA biosensor based on electrodeposited gold nanostructures as a transducer substrate combined with toluidine blue (TB) as a redox marker. Binding of TB with the single and double stranded DNA (ssDNA and dsDNA) was shortly investigated, and based on the results, TB could discriminate between ssDNA and dsDNA. A specific thiolated ssDNA sequence was immobilized on the transducer substrate, and DNA hybridization was followed by differential pulse voltammetry. The DNA biosensor showed excellent performances with high sensitivity and good selectivity. The DNA biosensor was applied to detect a synthetic target in a linear range of 1.0 × 10-1.0 × 10 mol L with a limit of detection (LOD) of 4.7 × 10 mol L. In addition, LOD of the DNA biosensor for the detection of genomic DNA was found to be 30.1 ng μL.

摘要

细菌、寄生虫和病毒广泛存在于环境中,是潜在的病原体,并可能引发感染。因此,需要灵敏、快速的方法来鉴定病原体,以提高生活质量。粪肠球菌通常定植于人体并威胁人类健康。本研究中,我们展示了一种新型电化学 DNA 生物传感器的构建,该传感器以电沉积金纳米结构作为传感器基底,结合甲苯胺蓝(TB)作为氧化还原标记物。我们对 TB 与单链 DNA(ssDNA)和双链 DNA(dsDNA)的结合短时间内进行了研究,并基于结果表明 TB 可以区分 ssDNA 和 dsDNA。经巯基化的 ssDNA 序列固定在传感器基底上,随后通过差分脉冲伏安法进行 DNA 杂交。该 DNA 生物传感器具有高灵敏度和良好的选择性。该 DNA 生物传感器用于检测合成靶标,线性范围为 1.0×10-1.0×10 mol L,检测限(LOD)为 4.7×10 mol L。此外,该 DNA 生物传感器用于检测基因组 DNA 的 LOD 为 30.1 ng μL。

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