An Electrochemical Immunosensor for Detection of Staphylococcus aureus Bacteria Based on Immobilization of Antibodies on Self-Assembled Monolayers-Functionalized Gold Electrode

被引:76
作者
Braiek, Mohamed [1 ,2 ]
Rokbani, Karima Bekir [3 ]
Chrouda, Amani [1 ,2 ]
Mrabet, Bechir [3 ]
Bakhrouf, Amina [3 ]
Maaref, Abderrazak [1 ]
Jaffrezic-Renault, Nicole [2 ]
机构
[1] Fac Sci Monastir, Lab Phys & Chim Interfaces, Ave Environm, Monastir 5019, Tunisia
[2] Univ Claude Bernard, Inst Sci Analyt, UMR CNRS 5280, F-69622 Villeurbanne, France
[3] Fac Pharm Monastir, Lab Anal Traitement & Valorisat Polluants IEnviro, Monastir, Tunisia
来源
BIOSENSORS-BASEL | 2012年 / 2卷 / 04期
关键词
Staphylococcus aureus; self-assembled monolayer; immunosensor; cyclic voltammetry; electrochemical impedance spectroscopy;
D O I
10.3390/bios2040417
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
The detection of pathogenic bacteria remains a challenge for the struggle against biological weapons, nosocomial diseases, and for food safety. In this research, our aim was to develop an easy-to-use electrochemical immunosensor for the detection of pathogenic Staphylococcus aureus ATCC25923. The biosensor was elaborated by the immobilization of anti-S. aureus antibodies using a self-assembled monolayer (SAMs) of 3-Mercaptopropionic acid (MPA). These molecular assemblies were spontaneously formed by the immersion of the substrate in an organic solvent containing the SAMs that can covalently bond to the gold surface. The functionalization of the immunosensor was characterized using two electrochemical techniques: cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). Here, the analysis was performed in phosphate buffer with ferro/ferricyanide as the redox probe. The EIS technique was used for affinity assays: antibody-cell binding. A linear relationship between the increment was observed between 10 and 106 CFU/mL. The limit of detection (LOD) was observed at 10 CFU/mL, and the reproducibility was calculated to 8%. Finally, a good selectivity versus E.coli and S. epidermidis was obtained for our developed immunosensor demonstrating its specificity towards only S.aureus.
引用
收藏
页码:417 / 426
页数:10
相关论文
共 16 条
[1]   Detection of pathogenic Staphylococcus aureus bacteria by gold based immunosensors [J].
Boujday, Souhir ;
Briandet, Romain ;
Salmain, Michele ;
Herry, Jean-Marie ;
Marnet, Pierre-Guy ;
Gautier, Michel ;
Pradier, Claire-Marie .
MICROCHIMICA ACTA, 2008, 163 (3-4) :203-209
[2]   Pathogenic micro-organisms in milk and dairy products: the situation in France and in Europe [J].
Brisabois, A ;
Lafarge, V ;
Brouillaud, A ;
de Buyser, ML ;
Collette, C ;
Garin-Bastuji, B ;
Thorel, MF .
REVUE SCIENTIFIQUE ET TECHNIQUE DE L OFFICE INTERNATIONAL DES EPIZOOTIES, 1997, 16 (02) :452-471
[3]  
Chaubeau-Duffour C., 1992, POINT VET, V24, P33
[4]  
De Buyser M.-L., 1994, POINT VET, V26, P79
[5]   Electrochemical immunosensor designs for the determination of Staphylococcus aureus using 3,3-dithiodipropionic acid di(N-succinimidyl ester)-modified gold electrodes [J].
Escamilla-Gomez, Vanessa ;
Campuzano, Susana ;
Pedrero, Maria ;
Pingarron, Jose M. .
TALANTA, 2008, 77 (02) :876-881
[6]   Dielectric spectroscopy in a micromachined flow cytometer: theoretical and practical considerations [J].
Gawad, S ;
Cheung, K ;
Seger, U ;
Bertsch, A ;
Renaud, P .
LAB ON A CHIP, 2004, 4 (03) :241-251
[7]   Electrochemical Impedance Spectroscopy (EIS) of ion sensors Direct modeling and inverse problem solving using the Nernst-Planck-Poisson (NPP) model and the HGS(FP) optimization strategy [J].
Grysakowski, B. ;
Jasielec, J. J. ;
Wierzba, B. ;
Sokalski, T. ;
Lewenstam, A. ;
Danielewski, M. .
JOURNAL OF ELECTROANALYTICAL CHEMISTRY, 2011, 662 (01) :143-149
[8]   Detection of Escherichia coli and Salmonella typhimurium using interdigitated microelectrode capacitive immunosensors:: The importance of transducer geometry [J].
Laczka, Olivier ;
Baldrich, Eva ;
Munoz, Francesc Xavier ;
del Campo, Fco Javier .
ANALYTICAL CHEMISTRY, 2008, 80 (19) :7239-7247
[9]   Label-free capacitive immunosensor based on quartz crystal Au electrode for rapid and sensitive detection of Escherichia coli O157:H7 [J].
Li, Dujuan ;
Feng, Yangyang ;
Zhou, Ling ;
Ye, Zunzhong ;
Wang, Jianping ;
Ying, Yibin ;
Ruan, Chuanmin ;
Wang, Ronghui ;
Li, Yanbin .
ANALYTICA CHIMICA ACTA, 2011, 687 (01) :89-96
[10]   Esterification of Self-Assembled Carboxylic-Acid-Terminated Thiol Monolayers in Acid Environment: A Time-Dependent Study [J].
Myrskog, Annica ;
Anderson, Henrik ;
Aastrup, Teodor ;
Ingemarsson, Bjorn ;
Liedberg, Bo .
LANGMUIR, 2010, 26 (02) :821-829