An electrochemical genosensor for Salmonella typhi on gold nanoparticles-mercaptosilane modified screen printed electrode

被引:50
作者
Das, Ritu [1 ]
Sharma, Mukesh K. [1 ]
Rao, Vepa K. [1 ]
Bhattacharya, B. K. [1 ]
Garga, Iti [1 ]
Venkatesh, V. [2 ]
Upadhyay, Sanjay [1 ]
机构
[1] Defence Res & Dev Estab, Gwalior 474002, India
[2] IIT, Dept Chem, Kanpur 208016, Uttar Pradesh, India
关键词
Biosensor; Differential pulse voltammetry; DNA; Mercaptopropyl trimethoxy silane; Screen printed electrode; SEQUENCE-SPECIFIC DETECTION; METHYLENE-BLUE; SENSITIVE DETECTION; RAPID DETECTION; DNA SENSOR; LABEL-FREE; PCR; TIME; IDENTIFICATION; AMPLIFICATION;
D O I
10.1016/j.jbiotec.2014.08.002
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
In this work, we fabricated a system of integrated self-assembled layer of organosilane 3-mercaptopropyltrimethoxy silane (MPTS) on the screen printed electrode (SPE) and electrochemically deposited gold nanoparticle for Salmonella typhi detection employing Vi gene as a molecular marker. Thiolated DNA probe was immobilized on a gold nanoparticle (AuNP) modified SPE for DNA hybridization assay using methylene blue as redox (electroactive) hybridization indicator, and signal was monitored by differential pulse voltammetry (DPV) method. The modified SPE was characterized by cyclic voltammetry(CV), electrochemical impedance spectroscopy (EIS), and atomic force microscopy (AFM) method. The DNA biosensor showed excellent performances with high sensitivity and good selectivity. The current response was linear with the target sequence concentrations ranging from 1.0 x 10(-11) to 0.5 x 10(-8) M and the detection limit was found to be 50 (+/- 2.1) pM. The DNA biosensor showed good discrimination ability to the one-base, two-base and three-base mismatched sequences. The fabricated genosensor could also be regenerated easily and reused for three to four times for further hybridization studies. (C) 2014 Elsevier B.V. All rights reserved.
引用
收藏
页码:9 / 16
页数:8
相关论文
共 50 条
[1]  
Alfonso A. S., 2013, BIOSENS BIOELECTRON, V40, P121
[2]   FORMATION OF MONOLAYERS BY THE COADSORPTION OF THIOLS ON GOLD - VARIATION IN THE HEAD GROUP, TAIL GROUP, AND SOLVENT [J].
BAIN, CD ;
EVALL, J ;
WHITESIDES, GM .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1989, 111 (18) :7155-7164
[3]   Distance-Dependent Electron Transfer at Passivated Electrodes Decorated by Gold Nanoparticles [J].
Barfidokht, Abbas ;
Ciampi, Simone ;
Luais, Erwann ;
Darwish, Nadim ;
Gooding, J. Justin .
ANALYTICAL CHEMISTRY, 2013, 85 (02) :1073-1080
[4]   Typhoid and paratyphoid fever [J].
Bhan, MK ;
Bahl, R ;
Bhatnagar, S .
LANCET, 2005, 366 (9487) :749-762
[5]  
Brad AJ., 2000, Electrochemical Methods: Fundamentals and Applications
[6]   Molecular beacon-metal nanowire interface: Effect of probe sequence and surface coverage on sensor performance [J].
Cederquist, Kristin B. ;
Golightly, Rebecca Stoermer ;
Keating, Christine D. .
LANGMUIR, 2008, 24 (16) :9162-9171
[7]  
Chaicumpa W, 1995, ASIAN PAC J ALLERGY, V13, P159
[8]   5′-thiolated oligonucleotides on (3-mercaptopropyl)trimethoxysilane-mica:: Surface topography and coverage [J].
Cheung, MKL ;
Trau, D ;
Yeung, KL ;
Carles, M ;
Sucher, NJ .
LANGMUIR, 2003, 19 (14) :5846-5850
[9]   A Polymer-Based Electrochemical DNA Biosensor for Salmonella: Preparation, Characterization and Calibration [J].
Diaz-Serrano, Madeline ;
Rosado, Arelys ;
del Pilar, Joselyn ;
Arias, Melvin ;
Guadalupe, Ana R. .
ELECTROANALYSIS, 2011, 23 (08) :1830-1841
[10]   Rapid Classification and Identification of Salmonellae at the Species and Subspecies Levels by Whole-Cell Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry [J].
Dieckmann, Ralf ;
Helmuth, Reiner ;
Erhard, Marcel ;
Malorny, Burkhard .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2008, 74 (24) :7767-7778