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壳聚糖-氧化铁纳米复合材料平台用于淋病奈瑟菌检测的错配识别 DNA 杂交,该菌可引起性传播疾病。

Chitosan-iron oxide nano-composite platform for mismatch-discriminating DNA hybridization for Neisseria gonorrhoeae detection causing sexually transmitted disease.

机构信息

DST Centre on Biomolecular Electronics, Biomedical Instrumentation Section, National Physical Laboratory CSIR, Dr KS Krishnan Marg, New Delhi 110012, India.

出版信息

Biosens Bioelectron. 2011 Feb 15;26(6):2967-74. doi: 10.1016/j.bios.2010.11.047. Epub 2010 Dec 7.

Abstract

Electrochemically fabricated nano-composite film of chitosan (CH)-iron oxide (Fe(3)O(4)) has been used to detect gonorrhoea, a sexually transmitted disease (STD) via immobilization of biotinylated probe DNA (BDNA) using avidin-biotin coupling for rapid and specific (mismatch-discriminating) DNA hybridization. The presence of Fe(3)O(4) nanoparticles (∼18nm) increases the electro-active surface area of the nano-biocomposite that provides desirable environment for loading of DNA with better conformation leading to increased electron transfer kinetics between the medium and electrode. The differential pulse voltammetric (DPV) studies have been conducted using BDNA/avidin/CH-Fe(3)O(4)/ITO electrode owing to the reduction of the methylene blue (MB) indicator and investigate electron transfer between MB moieties and electrode for one and two-bases mismatch. This STD biosensor is found to have a detection limit (1 × 10(-15)M) and a wide dynamic range (from 1 × 10(-16)M to 1 × 10(-6)M) using the complementary target DNA. In addition, the sensing system can be utilized to accurately discriminate complementary sequence from mismatch sequences.

摘要

电化学制备的壳聚糖 (CH)-氧化铁 (Fe(3)O(4)) 纳米复合材料膜已被用于通过生物素化探针 DNA (BDNA) 的固定化来检测淋病,淋病是一种性传播疾病 (STD),使用亲和素-生物素偶联进行快速和特异性 (错配区分) DNA 杂交。Fe(3)O(4)纳米粒子 (∼18nm) 的存在增加了纳米生物复合材料的电活性表面积,为 DNA 提供了更好构象的加载环境,从而导致介质和电极之间的电子转移动力学增加。由于亚甲蓝 (MB) 指示剂的还原,使用 BDNA/亲和素/CH-Fe(3)O(4)/ITO 电极进行了差分脉冲伏安法 (DPV) 研究,并研究了 MB 部分和电极之间的电子转移对于单碱基和双碱基错配。使用互补靶 DNA,该 STD 生物传感器具有检测限 (1 × 10(-15)M) 和宽动态范围 (从 1 × 10(-16)M 到 1 × 10(-6)M)。此外,该传感系统可用于准确区分互补序列和错配序列。

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