Immobilization of single stranded DNA probe onto polypyrrole-polyvinyl sulfonate for application to DNA hybridization biosensor

被引:49
作者
Arora, Kavita
Prabhakar, Nirmal
Chand, Subhash
Malhotra, B. D.
机构
[1] Natl Phys Lab, Biomol Elect & Conducting Polymer Res Grp, New Delhi 110012, India
[2] Indian Inst Technol, Dept Biochem Engn & Biotechnol, New Delhi 110012, India
来源
SENSORS AND ACTUATORS B-CHEMICAL | 2007年 / 126卷 / 02期
关键词
DNA biosensor; polypyrrole-polyvinyl sulfonate; biotin-avidin; hybridization;
D O I
10.1016/j.snb.2007.04.029
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Electrochemical DNA biosensor based on polypyrrole-polyvinyl sulfonate (PPy-PVS) coated onto Pt disc electrode (similar to 0.03 cm(2)) has been fabricated by immobilizing 25 bases long single stranded probe polydeoxycytidine (dC) using both biotin-avidin binding (indirect immobilization) and carbodiimide coupling (direct immobilization), to detect 25 bases long complementary target sequence polydeoxyguanosine (dG). These DNA immobilized electrodes have been characterized using differential pulse voltarnmetry (DPV), Fourier transform infrared spectroscopy (FT-IR) and scanning electron microscopy (SEM) techniques, respectively. Compared to the indirect method of immobilization, direct immobilization of dC probe results in enhanced detection limit by 2200 times (upto 0.01 fmol) and improved sensitivity by about six times. These DNA electrodes have response time of 30 s and are stable for about 4 months when stored at 4 degrees C. (C) 2007 Elsevier B.V. All rights reserved.
引用
收藏
页码:655 / 663
页数:9
相关论文
共 42 条
[11]   Novel hybridization indicator methylene blue for the electrochemical detection of short DNA sequences related to the hepatitis B virus [J].
Erdem, A ;
Kerman, K ;
Meric, B ;
Akarca, US ;
Ozsoz, M .
ANALYTICA CHIMICA ACTA, 2000, 422 (02) :139-149
[12]   Potentiometric study of polyaniline film synthesized with various dopants and composite-dopant: A comparative study [J].
Gaikwad, P. D. ;
Shirale, D. J. ;
Gade, V. K. ;
Savale, P. A. ;
Kakde, K. P. ;
Kharat, H. J. ;
Shirsat, M. D. .
BULLETIN OF MATERIALS SCIENCE, 2006, 29 (04) :417-420
[13]   Coimmobilization of urease and glutamate dehydrogenase in electrochemically prepared polypyrrole-polyvinyl sulfonate films [J].
Gambhir, A ;
Gerard, M ;
Mulchandani, AK ;
Malhotra, BD .
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY, 2001, 96 (1-3) :249-257
[14]   Electrochemical molecular analysis without nucleic acid amplification [J].
Gau, V ;
Ma, SC ;
Wang, H ;
Tsukuda, J ;
Kibler, J ;
Haake, DA .
METHODS, 2005, 37 (01) :73-83
[15]   A comparison of redox processes for polypyrrole/dodecylsulfate films in aqueous and non-aqueous media [J].
Hallik, A ;
Alumaa, A ;
Sammelselg, V ;
Tamm, J .
JOURNAL OF SOLID STATE ELECTROCHEMISTRY, 2001, 5 (04) :265-273
[16]   Recognition and detection of oligonucleotides in the presence of chromosomal DNA based on entrapment within conducting-polymer networks [J].
Jiang, M ;
Wang, J .
JOURNAL OF ELECTROANALYTICAL CHEMISTRY, 2001, 500 (1-2) :584-589
[17]   Interaction of antioxidant flavonoids with tRNA: Intercalation or external binding and comparison with flavonoid-DNA adducts [J].
Kanakis, CD ;
Tarantilis, PA ;
Polissiou, MG ;
Tajmir-Riahi, HA .
DNA AND CELL BIOLOGY, 2006, 25 (02) :116-123
[18]   Label-free electrochemical detection of DNA hybridization on gold electrode [J].
Kerman, K ;
Morita, Y ;
Takamura, Y ;
Tamiya, E .
ELECTROCHEMISTRY COMMUNICATIONS, 2003, 5 (10) :887-891
[19]   Voltammetric determination of DNA hybridization using methylene blue and self-assembled alkanethiol monolayer on gold electrodes [J].
Kerman, K ;
Ozkan, D ;
Kara, P ;
Meric, B ;
Gooding, JJ ;
Ozsoz, M .
ANALYTICA CHIMICA ACTA, 2002, 462 (01) :39-47
[20]   Poly(pyrrole) versus poly (3,4-ethylenedioxythiophene): implications for biosensor applications [J].
Kros, A ;
Sommerdijk, NAJM ;
Nolte, RJM .
SENSORS AND ACTUATORS B-CHEMICAL, 2005, 106 (01) :289-295