Maalouf Rita, Fournier-Wirth Chantal, Coste Joliette, Chebib Hanna, Saïkali Youssef, Vittori Olivier, Errachid Abdelhamid, Cloarec Jean-Pierre, Martelet Claude, Jaffrezic-Renault Nicole
AMPERE, UMR/CNRS 5005 and INL, UMR/CNRS 5270, Ecole Centrale de Lyon, 36, 69134 Ecully, France.
Anal Chem. 2007 Jul 1;79(13):4879-86. doi: 10.1021/ac070085n. Epub 2007 May 25.
The low but known risk of bacterial contamination has emerged as the greatest residual threat of transfusion-transmitted diseases. Label-free detection of a bacterial model, Escherichia coli, is performed using nonfaradic electrochemical impedance spectroscopy (EIS). Biotinylated polyclonal anti-E. coli is linked to a mixed self-assembled monolayer (SAM) on a gold electrode through a strong biotin-neutravidin interaction. The binding of one antibody molecule for 3.6 neutravidin molecules is determined using the surface plasmon resonance (SPR). The detection limit of E. coli found by SPR is 10(7) cfu/mL. After modeling the impedance Nyquist plot of E. coli/anti-E. coli/mixed SAM/gold electrode for increasing concentrations of E. coli (whole bacteria or lysed bacteria), the main parameter that is modified is the polarization resistance RP. A sigmoid variation of RP is observed when the log concentration of bacteria (whole or lysed) increases. A concentration of 10 cfu/mL whole bacteria is detected by EIS measurements while 103 cfu/mL is detected for lysed E. coli.
细菌污染风险虽低但已知,已成为输血传播疾病最大的残留威胁。使用非法拉第电化学阻抗谱(EIS)对细菌模型大肠杆菌进行无标记检测。生物素化的抗大肠杆菌多克隆抗体通过生物素与中性抗生物素蛋白的强相互作用连接到金电极上的混合自组装单层(SAM)。使用表面等离子体共振(SPR)确定一个抗体分子与3.6个中性抗生物素蛋白分子的结合。通过SPR发现大肠杆菌的检测限为10(7) cfu/mL。在对不同浓度大肠杆菌(完整细菌或裂解细菌)的大肠杆菌/抗大肠杆菌/混合SAM/金电极的阻抗奈奎斯特图进行建模后,主要改变的参数是极化电阻RP。当细菌(完整或裂解)的对数浓度增加时,观察到RP呈S形变化。通过EIS测量检测到完整细菌的浓度为10 cfu/mL,而裂解大肠杆菌的检测浓度为103 cfu/mL。