Abstract
Non-specific absorption of enzyme and nucleotide is a main source leading to the background staining in enzyme-amplified amperometric detection of DNA. To eliminate the background staining in this system, the blocking procedure by mercapto-hexanol ( MCH) and bovine serum albumin ( BSA) was studied systematically. The results show that the anti-fouling effect of BSA is better than that of MCH. A simple and efficient blocking strategy employing BSA as the sole blocking reagent was established and applied in the detection of PMLRARα fusion gene in acute promyelocytic leukemia. It can effectively eliminate the background staining,shorten the pre-processing time,and achieve strong signal amplification.
Keywords
electrochemical DNA sensor, enzyme-amplified amperometric assay, bovine serum albumin, blocking
Publication Date
2010-05-28
Online Available Date
2010-05-28
Revised Date
2010-05-28
Received Date
2010-05-28
Recommended Citation
Miao HE, Wei CHEN, Xiong-wei XU, Ai-lin LIU, Xin-hua LIN.
Blocking Method in Enzyme-Amplified Amperometric DNA Biosensor[J]. Journal of Electrochemistry,
2010
,
16(2): Article 21.
DOI: 10.61558/2993-074X.3349
Available at:
https://jelectrochem.xmu.edu.cn/journal/vol16/iss2/21
References
[1]Caruso F.Nanoengineering of particle surfaces[J].Adv Mater,2001,13:11.
[2]Storhoff J J,Mirkin C A.Programmed materials synthe-sis with DNA[J].Chem Rev,1999,99:1849.
[3]Willner I,Willner B.Functional nanoparticle architec-tures for sensoric,optoelectronic,and bioelectronic ap-plications[J].Pure Appl Chem,2002,74:1773.
[4]Campbell C N,Gal D,Cristler N,et al.Enzyme ampli-fied amperometric sandwich test for RNA and DNA[J].Anal Chem,2002,74:158.
[5]Aguilar Z P,Fritsch I.Immobilized enzyme-linked DNA-hybridization assay with electrochemical detection for Cryptosporidium parvum hsp70mRNA[J].Anal Chem,2003,75:3890.
[6]Patolsky F,Lichtenstein A,Willner I.Detection of sin-gle-base DNA mutations by enzyme-amplified electronic transduction[J].Nat Biotechnol,2001,19:253.
[7]Zhang Jiong,Lao Ruojun,Song Shiping,et al.Design of an oligonucleotide-incorporated nonfouling surface andits application in electrochemical DNA sensors for highly sensitive and sequence-specific detection of target DNA[J].Anal Chem,2008,80(23):9029-9033.
[8]Wang Baozhen(王保珍),Du Xiaoyan(杜晓燕),Zheng Jing(郑晶),et al.Electrochemical sensor based on im-mobilization of single stranded deoxyribonucleic acid on Pt electrode surface by avidin-biotin system[J].Chinese Journal of Analytical Chemistry(分析化学研究报告),2005,33(6):789-792.
[9]Poirier G E,Pylant E D.The self-assembly mechanism of alkanethiols on Au(111)[J].Science,1996,272:1145-1148.
[10]Kai E,Sawata S,Ikebukuro K,et al.Detection of PCR products in solution using surface plasmon resonance[J].Analytical Chemistry,1999,71:796-800.
[11]Li Meisheng(李梅生),Li Weixing(李卫星),Xing Weihong(邢卫红),et al.Adsorption of bovine serum albumin on three kinds of ceramic membrane materials[J].Membrane Science and Technology(膜科学与技术),2007,27(5):18-21.
[12]Wan Ying,Zhang Jiong,Liu Gang,et al.Ligase-based multiple DNA analysis by using an electrochemical sensor array[J].Biosensors and bioelectronics,2009,24(5):1209-1212.
[13]Mao Xun,Jiang Jianhui,Xu Xiangmin,et al.Enzymatic amplification detection of DNA based on″molecular beacon″biosensors[J].Biosensors and Bioelectronics,2008,23:1555-1561.
[14]Levicky R,Herne T M,Tarlov M J,et al.Using self-as-sembly to control the structure of DNA monolayers on gold:A neutron reflectivity study[J].J Am Chem Soc,1998,120:9787-9792.