Suppr超能文献

动物组织中基因组DNA的高灵敏度安培检测。

Highly sensitive amperometric detection of genomic DNA in animal tissues.

作者信息

Xie Hong, Yu Yuan Hong, Mao Pei-Lin, Gao Zhiqiang

机构信息

Institute of Bioengineering and Nanotechnology, 51 Science Park Road, Singapore 117586, Republic of Singapore.

出版信息

Nucleic Acids Res. 2004 Jan 20;32(2):e15. doi: 10.1093/nar/gnh016.

DOI:10.1093/nar/gnh016
PMID:14734816
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC373372/
Abstract

A simple and highly sensitive method for the detection of genomic DNA in tissue samples is described. It is based on amperometric detection of target DNA by forming an analyte/polymeric activator bilayer on a gold electrode. The biotinylated target DNA is hybridized to oligonucleotide capture probes immobilized on the gold electrode, forming the first layer. A subsequent binding of glucose oxidase-avidin conjugate to the target DNA and the introduction of a second layer of a redox polymer to the electrode, via layer-by-layer electrostatic self-assembly, allow for electrochemical detection of the catalytic oxidation current of glucose in a PBS solution. Less than 2.0 fg of rat genomic DNA, for both regulated and house-keeping genes, can be easily detected in 2.5 microl droplets. The proposed procedure shows very high specificity for genomic DNA in a RT-PCR mixture.

摘要

本文描述了一种用于检测组织样本中基因组DNA的简单且高度灵敏的方法。该方法基于在金电极上形成分析物/聚合物激活剂双层来对目标DNA进行安培检测。生物素化的目标DNA与固定在金电极上的寡核苷酸捕获探针杂交，形成第一层。随后，葡萄糖氧化酶-抗生物素蛋白结合物与目标DNA结合，并通过逐层静电自组装将氧化还原聚合物的第二层引入电极，从而能够在PBS溶液中对葡萄糖的催化氧化电流进行电化学检测。在2.5微升液滴中，无论是调控基因还是管家基因，都能轻松检测到少于2.0 fg的大鼠基因组DNA。所提出的方法对RT-PCR混合物中的基因组DNA具有非常高的特异性。

相似文献

1
Highly sensitive amperometric detection of genomic DNA in animal tissues.动物组织中基因组DNA的高灵敏度安培检测。
Nucleic Acids Res. 2004 Jan 20;32(2):e15. doi: 10.1093/nar/gnh016.
2
Amperometric detection of nucleic acid at femtomolar levels with a nucleic acid/electrochemical activator bilayer on gold electrode.利用核酸/电化学活化剂双层在金电极上对飞摩尔水平的核酸进行安培检测。
Anal Chem. 2004 Mar 15;76(6):1611-7. doi: 10.1021/ac0350965.
3
Enzyme-amplified amperometric sandwich test for RNA and DNA.用于RNA和DNA的酶放大安培夹心试验。
Anal Chem. 2002 Jan 1;74(1):158-62. doi: 10.1021/ac015602v.
4
Redox polymer and probe DNA tethered to gold electrodes for enzyme-amplified amperometric detection of DNA hybridization.连接到金电极上的氧化还原聚合物和探针DNA用于酶放大安培法检测DNA杂交。
Anal Chem. 2006 Apr 15;78(8):2710-6. doi: 10.1021/ac0521100.
5
A nucleic acid biosensor for gene expression analysis in nanograms of mRNA.
Anal Chem. 2004 Jul 15;76(14):4023-9. doi: 10.1021/ac049839d.
6
Enzyme-amplified electrochemical detection of DNA using electrocatalysis of ferrocenyl-tethered dendrimer.使用二茂铁连接的树枝状大分子的电催化对DNA进行酶放大电化学检测。
Anal Chem. 2003 Nov 1;75(21):5665-72. doi: 10.1021/ac034253x.
7
Highly sensitive detection of target gene by electrochemical method.
Nucleic Acids Symp Ser. 1999(42):185-6. doi: 10.1093/nass/42.1.185.
8
Electrogenerated chemiluminescence. 72. Determination of immobilized DNA and C-reactive protein on Au(111) electrodes using tris(2,2'-bipyridyl)ruthenium(II) labels.电致化学发光。72. 使用三联吡啶钌（II）标记物测定金（111）电极上固定的DNA和C反应蛋白。
Anal Chem. 2003 Nov 1;75(21):5825-34. doi: 10.1021/ac034596v.
9
Enzyme-amplified amperometric detection of 3000 copies of DNA in a 10-microL droplet at 0.5 fM concentration.在10微升液滴中，以0.5飞摩尔浓度对3000个DNA拷贝进行酶放大安培检测。
Anal Chem. 2003 Jul 1;75(13):3267-9. doi: 10.1021/ac034445s.
10
An ultrasensitive protein array based on electrochemical enzyme immunoassay.
Front Biosci. 2005 May 1;10:1654-60. doi: 10.2741/1649.

引用本文的文献

1
Assembling Amperometric Biosensors for Clinical Diagnostics.用于临床诊断的安培型生物传感器组装
Sensors (Basel). 2008 Feb 27;8(3):1366-1399. doi: 10.3390/s8031366.
2
DNA-based bioanalytical microsystems for handheld device applications.用于手持设备应用的基于DNA的生物分析微系统。
Anal Chim Acta. 2006 Jan 18;556(1):26-37. doi: 10.1016/j.aca.2005.05.075. Epub 2005 Jul 7.

本文引用的文献

1
Electronic Transduction of Polymerase or Reverse Transcriptase Induced Replication Processes on Surfaces: Highly Sensitive and Specific Detection of Viral Genomes.
Angew Chem Int Ed Engl. 2001 Jun 18;40(12):2261-2265. doi: 10.1002/1521-3773(20010618)40:12<2261::AID-ANIE2261>3.0.CO;2-P.
2
Surface Plasmon Resonance Imaging Measurements of Electrostatic Biopolymer Adsorption onto Chemically Modified Gold Surfaces.静电生物聚合物吸附到化学修饰金表面的表面等离子体共振成像测量
Anal Chem. 1997 Apr 1;69(7):1449-56. doi: 10.1021/ac961012z.
3
Amperometric detection of nucleic acid at femtomolar levels with a nucleic acid/electrochemical activator bilayer on gold electrode.利用核酸/电化学活化剂双层在金电极上对飞摩尔水平的核酸进行安培检测。
Anal Chem. 2004 Mar 15;76(6):1611-7. doi: 10.1021/ac0350965.
4
Enzyme-amplified amperometric detection of 3000 copies of DNA in a 10-microL droplet at 0.5 fM concentration.在10微升液滴中，以0.5飞摩尔浓度对3000个DNA拷贝进行酶放大安培检测。
Anal Chem. 2003 Jul 1;75(13):3267-9. doi: 10.1021/ac034445s.
5
Electrodeposition of redox polymers and co-electrodeposition of enzymes by coordinative crosslinking.
Angew Chem Int Ed Engl. 2002 Mar 1;41(5):810-3. doi: 10.1002/1521-3773(20020301)41:5<810::aid-anie810>3.0.co;2-i.
6
Detection of single-base DNA mutations by enzyme-amplified electronic transduction.通过酶扩增电子转导检测单碱基DNA突变
Nat Biotechnol. 2001 Mar;19(3):253-7. doi: 10.1038/85704.
7
Mutation detection by electrocatalysis at DNA-modified electrodes.通过DNA修饰电极上的电催化进行突变检测。
Nat Biotechnol. 2000 Oct;18(10):1096-100. doi: 10.1038/80301.
8
From DNA biosensors to gene chips.从DNA生物传感器到基因芯片。
Nucleic Acids Res. 2000 Aug 15;28(16):3011-6. doi: 10.1093/nar/28.16.3011.
9
Single-base mismatch detection based on charge transduction through DNA.基于通过DNA进行电荷传导的单碱基错配检测。
Nucleic Acids Res. 1999 Dec 15;27(24):4830-7. doi: 10.1093/nar/27.24.4830.
10
Electrochemical quantitation of DNA immobilized on gold.固定在金上的DNA的电化学定量分析。
Anal Chem. 1998 Nov 15;70(22):4670-7. doi: 10.1021/ac980037q.

文献检索

告别复杂PubMed语法，用中文像聊天一样搜索，搜遍4000万医学文献。AI智能推荐，让科研检索更轻松。

立即免费搜索

文件翻译

保留排版，准确专业，支持PDF/Word/PPT等文件格式，支持 12+语言互译。

免费翻译文档

深度研究

AI帮你快速写综述，25分钟生成高质量综述，智能提取关键信息，辅助科研写作。

立即免费体验