Sensitive detection of lip genes by electrochemical DNA sensor and its application in polymerase chain reaction amplicons from Phanerochaete chrysosporium

被引：52

作者：

Tang, Lin ^{[1
]}

Zeng, Guangming ^{[1
]}

Shen, Guoli ^{[2
]}

Li, Yuanping ^{[1
]}

Liu, Can ^{[1
]}

Li, Zhen ^{[1
]}

Luo, Jie ^{[1
]}

Fan, Changzheng ^{[1
]}

Yang, Chunping ^{[1
]}

机构：

[1] Hunan Univ, Coll Environm Sci & Engn, Changsha 410082, Hunan, Peoples R China

[2] Hunan Univ, State Key Lab Chemo Biosensing & Chemometr, Coll Chem & Chem Engn, Changsha 410082, Hunan, Peoples R China

来源：

BIOSENSORS & BIOELECTRONICS | 2009年 / 24卷 / 05期

基金：

中国国家自然科学基金;

关键词：

lip genes; Electrochemical DNA sensor; Polymerase chain reaction; Phanerochaete chrysosporium; LIGNIN PEROXIDASE; AMPEROMETRIC DETECTION; NUCLEIC-ACID; MONOLAYERS; IMMOBILIZATION; HYBRIDIZATION; GENOSENSOR; BILAYER;

D O I：

10.1016/j.bios.2008.09.031

中图分类号：

Q6 [生物物理学];

学科分类号：

071011 ;

摘要：

An electrochemical DNA sensor based on the sandwich hybridization recognition of target sequence of lignin peroxidase (lip) genes on a gold electrode was developed. A monolayer of thiolated capture probe was formed on a gold electrode through self-assembling. Following hybridizations with target nucleic acid and biotinylated detection probe, streptavidin-horseradish peroxidase (HRP) conjugate was applied to the electrode. The DNA conformation and surface coverage on electrode were characterized by impedance spectroscopy and square wave voltammetry. The experimental variables were optimized to maximize the hybridization efficiency, detection sensitivity and speed up the assay time. The amperometric current response to HRP-catalyzed reaction was linearly related to the natural logarithm of the target nucleic acid concentration in the range from 0.6 to 30 nM, with the correlation coefficient of 0.9722. The detection limit was 0.03 nM. Synthesized oligonucleotide as well as Phanerochaete chrysosporium lip gene fragments amplified using polymerase chain reaction and digested by restriction endonucleases were tested. The DNA sensor exhibited good precision, stability, sensitivity, and selectivity, and discriminated satisfactorily against mismatched nucleic acid samples of similar lengths. (C) 2008 Elsevier B.V. All rights reserved.

引用

页码：1474 / 1479

页数：6

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