QCM immunosensor with nanoparticle amplification for detection of Escherichia coli O157:H7

A quartz crystal microbalance (QCM) immunosensor was developed and evaluated for detection of Escherichia coli O157:H7. The immunosensor was fabricated by self-assembling of protein A and affinity-purified anti-E. coli O157:H7 antibodies on the gold electrode of an AT-cut piezoelectric quartz crystal. To enhance the sensitivity of the QCM immunosensor, nanoparticle-antibody conjugates, which were prepared using streptavidin-conjugated nanoparticles (145 nm diameter) and biotinylated anti-E. coli antibodies, were used for signal amplification. After the binding of E. coli O157:H7 cells with the antibodies immobilized on the electrode, nanoparticle-antibody conjugates were introduced as mass amplifiers. Compared to the direct detection of E. coli O157:H7, the binding of the nanoparticle conjugates further resulted in a decrease in resonant frequency and an increase in resonant resistance, and the detection sensitivity was improved by five orders of magnitude by lowering the detection limit from 107 to 102 CFU/mL. The sensor specificity and nonspecific adsorption of nanoparticle-antibody conjugates were also investigated.

[1]  Xiao-li Su,et al.  SURFACE PLASMON RESONANCE AND QUARTZ CRYSTAL MICROBALANCE IMMUNOSENSORS FOR DETECTION OF ESCHERICHIA COLI O157:H7 , 2005 .

[2]  Antje J Baeumner,et al.  Biosensors for environmental pollutants and food contaminants , 2003, Analytical and bioanalytical chemistry.

[3]  A. Deisingh,et al.  Strategies for the detection of Escherichia coli O157:H7 in foods , 2004, Journal of applied microbiology.

[4]  R. Buchanan,et al.  Foodborne Disease Significance of Escherichia coli O157:H7 and Other Enterohemorrhagic E. coli , 1997 .

[5]  Dmitri Ivnitski,et al.  Biosensors for detection of pathogenic bacteria , 1999 .

[6]  Egon Matijević,et al.  Fine Particles: Science and Technology , 1989 .

[7]  I. Park,et al.  Operational characteristics of an antibody-immobilized QCM system detecting Salmonella spp. , 2000, Biosensors & bioelectronics.

[8]  X. Mao,et al.  A nanoparticle amplification based quartz crystal microbalance DNA sensor for detection of Escherichia coli O157:H7. , 2006, Biosensors & bioelectronics.

[9]  R. Tauxe,et al.  The epidemiology of infections caused by Escherichia coli O157:H7, other enterohemorrhagic E. coli, and the associated hemolytic uremic syndrome. , 1991, Epidemiologic reviews.

[10]  Yanbin Li,et al.  A self-assembled monolayer-based piezoelectric immunosensor for rapid detection of Escherichia coli O157:H7. , 2004, Biosensors & bioelectronics.

[11]  Michael Thompson,et al.  Detection of infectious and toxigenic bacteria. , 2002, The Analyst.

[12]  Yanbin Li,et al.  A QCM immunosensor for Salmonella detection with simultaneous measurements of resonant frequency and motional resistance. , 2005, Biosensors & bioelectronics.

[13]  Lauro T. Kubota,et al.  Review of the use of biosensors as analytical tools in the food and drink industries , 2002 .

[14]  A. Deisingh,et al.  Biosensors for the detection of bacteria. , 2004, Canadian journal of microbiology.

[15]  K. Marx,et al.  The Quartz Crystal Microbalance as a Continuous Monitoring Tool for the Study of Endothelial Cell Surface Attachment and Growth , 2000, Biotechnology progress.

[16]  S. Yao,et al.  A Comparative Study on the Viscoelasticity and Morphology of Polyaniline Films Galvanostatically Grown on Bare and 4-Aminothiophenol-Modified Gold Electrodes Using an Electrochemical Quartz Crystal Impedance System and SEM , 2001, Analytical sciences : the international journal of the Japan Society for Analytical Chemistry.

[17]  Y. Fung,et al.  Self-assembled monolayers as the coating in a quartz piezoelectric crystal immunosensor to detect Salmonella in aqueous solution. , 2001, Analytical chemistry.