Detection of Salmonella typhimurium using an electrochemical immunosensor.

An electrochemical immunosensor based on screen-printed gold working electrode with onboard carbon counter and silver-silver chloride pseudo-reference electrode for Salmonella typhimurium detection is described in this paper. Monoclonal anti-S. typhimurium antibody was immobilized using physical and covalent immobilization via amine coupling of carboxymethyldextran on the surface of the gold working electrode. A direct sandwich enzyme-linked immunosorbent assays (ELISA) format was then developed and optimized using a polyclonal anti-Salmonella antibodies conjugated to horseradish peroxidase (HRP) as the enzyme label. 3,3',5,5'-Tetramethylbenzidine dihydrochloride (TMB)/H(2)O(2) was used as the enzyme mediator/substrate system. Electrochemical detection was conducted using chronoamperometry at -200 mV vs. onboard screen-printed Ag-AgCl pseudo-reference electrode. The applied potential was selected through the study of the electrochemical behaviour of bare gold electrode with TMB-H(2)O(2)-IgG-HRP system. S. typhimurium detection of 5x10(3) cells ml(-1) and approximately 20 cells ml(-1) was achieved respectively for physical and covalent antibody immobilization. The developed sensor was then compared to a commercial ELISA kit and a chromogenic agar plating method for meat samples analysis. The sensor format shows a promising technology for simple and sensitive detection system for Salmonella contamination. Rapid detection of Salmonella is a key to the prevention and identification of problems related to health and safety.

[1]  J. Schlundt,et al.  New directions in foodborne disease prevention. , 2002, International journal of food microbiology.

[2]  R. O'Kennedy,et al.  Advances in biosensors for detection of pathogens in food and water , 2003 .

[3]  Ibtisam E. Tothill Rapid and on-line instrumentation for food quality assurance. , 2003 .

[4]  M. Mozola Genetics-based methods for detection of Salmonella spp. in foods. , 2006, Journal of AOAC International.

[5]  D. Swerdlow,et al.  Precautions against biological and chemical terrorism directed at food and water supplies. , 2001, Public health reports.

[6]  D. O'Shannessy,et al.  Immobilization chemistries suitable for use in the BIAcore surface plasmon resonance detector. , 1992, Analytical biochemistry.

[7]  G. Palleschi,et al.  Development of SYBR‐Green Real‐Time PCR and a Multichannel Electrochemical Immunosensor for Specific Detection of Salmonella enterica , 2006 .

[8]  M. Rapp,et al.  Covalent bound sensing layers on surface acoustic wave (SAW) biosensors. , 2001, Biosensors & bioelectronics.

[9]  Liju Yang,et al.  Electrical impedance spectroscopy for detection of bacterial cells in suspensions using interdigitated microelectrodes. , 2008, Talanta.

[10]  Olivier Lazcka,et al.  Pathogen detection: a perspective of traditional methods and biosensors. , 2007, Biosensors & bioelectronics.

[11]  John Gray,et al.  Water contamination emergencies: enhancing our response. , 2006 .

[12]  Giuseppe Palleschi,et al.  3,3′,5,5′-Tetramethylbenzidine as electrochemical substrate for horseradish peroxidase based enzyme immunoassays. A comparative study , 1998 .

[13]  Paul Finglas,et al.  Encyclopedia of food sciences and nutrition , 2003 .

[14]  L. McCaig,et al.  Food-related illness and death in the United States. , 1999, Emerging infectious diseases.

[15]  Bo Johnsson,et al.  Comparison of methods for immobilization to carboxymethyl dextran sensor surfaces by analysis of the specific activity of monoclonal antibodies , 1995, Journal of molecular recognition : JMR.

[16]  E. Tondo,et al.  Evaluation of an indirect ELISA for the detection of Salmonella in chicken meat , 2006 .

[17]  E. Alocilja,et al.  Market analysis of biosensors for food safety. , 2003, Biosensors & bioelectronics.

[18]  G. Volpe,et al.  A RAPID ELECTROCHEMICAL ELISA FOR THE DETECTION OF SALMONELLA IN MEAT SAMPLES , 2001 .

[19]  I. Tothill,et al.  Development and characterisation of disposable gold electrodes, and their use for lead(II) analysis , 2006, Analytical and bioanalytical chemistry.

[20]  W H Andrews,et al.  Relative effectiveness of selenite cystine broth, tetrathionate broth, and rappaport-vassiliadis medium for recovery of Salmonella spp. from raw flesh, highly contaminated foods, and poultry feed: collaborative study. , 1996, Journal of AOAC International.