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用于痕量DNA定量的电容式生物传感器。

Capacitive biosensor for quantification of trace amounts of DNA.

作者信息

Numnuam Apon, Kanatharana Proespichaya, Mattiasson Bo, Asawatreratanakul Punnee, Wongkittisuksa Booncharoen, Limsakul Chusak, Thavarungkul Panote

机构信息

Trace Analysis and Biosensor Research Center, Prince of Songkla University, Hat Yai, Songkhla 90112, Thailand.

出版信息

Biosens Bioelectron. 2009 Apr 15;24(8):2559-65. doi: 10.1016/j.bios.2009.01.005. Epub 2009 Jan 14.

Abstract

A flow injection capacitive biosensor system to detect trace amounts DNA has been developed based on the affinity binding between immobilized histone and DNA. Histones from calf thymus and shrimp were immobilized on gold electrodes covered with self-assembled monolayer (SAM) of thioctic acid. Each of these histones was used to detect DNA from calf thymus, shrimp and Escherichia coli. The studies indicated that histones can bind better with DNA from the same source and give higher sensitivity than the binding with DNA from different sources. Under optimum conditions, both histones from calf thymus and shrimp provided the same lower detection limit of 10(-5) ng l(-1) for DNA from different sources, i.e., calf thymus, shrimp and E. coli. The standard curve for the affinity reaction between calf thymus histone and DNA shows sigmoidal behavior and two linear ranges, 10(-5) to 10(-2) ng l(-1) and 10(-1) to 10(2) ng l(-1), could be obtained. The immobilized histones were stable and after regeneration good reproducibility of the signal could be obtained up to 43 times with a %R.S.D. of 3.1. When applied to analyze residual DNA in crude protein extracted from white shrimp recoveries were obtained between 80% and 116%.

摘要

基于固定化组蛋白与DNA之间的亲和结合,开发了一种用于检测痕量DNA的流动注射电容式生物传感器系统。将来自小牛胸腺和虾的组蛋白固定在覆盖有硫辛酸自组装单层(SAM)的金电极上。每种组蛋白都用于检测来自小牛胸腺、虾和大肠杆菌的DNA。研究表明,组蛋白与来自同一来源的DNA结合更好,并且比与来自不同来源的DNA结合具有更高的灵敏度。在最佳条件下,来自小牛胸腺和虾的组蛋白对来自不同来源(即小牛胸腺、虾和大肠杆菌)的DNA都提供了相同的10^(-5) ng l^(-1)的较低检测限。小牛胸腺组蛋白与DNA之间亲和反应的标准曲线呈S形,可获得两个线性范围,即10^(-5)至10^(-2) ng l^(-1)和10^(-1)至10^(2) ng l^(-1)。固定化组蛋白稳定,再生后信号具有良好的重现性,最多可重复43次,相对标准偏差为3.1%。当应用于分析从白虾中提取的粗蛋白中的残留DNA时,回收率在80%至116%之间。

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