Impedance biosensing using phages for bacteria detection: generation of dual signals as the clue for in-chip assay confirmation.

In the present work, we compare the use of antibodies (Ab) and phages as bioreceptors for bacteria biosensing by Electrochemical Impedance Spectroscopy (EIS). With this aim, both biocomponents have been immobilised in parallel onto interdigitated gold microelectrodes. The produced surfaces have been characterised by EIS and Fourier Transform Infra-Red (FTIR) Spectroscopy and have been applied to bacteria detection. Compared to immunocapture, detection using phages generates successive dual signals of opposite trend over time, which consist of an initial increase in impedance caused by bacteria capture followed by impedance decrease attributed to phage-induced lysis. Such dual signals can be easily distinguished from those caused by non-specific adsorption and/or crossbinding, which helps to circumvent one of the main drawbacks of reagentless biosensors based in a single target-binding event. The described strategy has generated specific detection of Escherichia coli in the range of 10(4)-10(7) CFU mL(-1) and minimal interference by non-target Lactobacillus. We propose that the utilisation of phages as capture biocomponent for bacteria capture and EIS detection allows in-chip signal confirmation.

[1]  C. Roychaudhuri,et al.  Macroporous silicon based simple and efficient trapping platform for electrical detection of Salmonella typhimurium pathogens. , 2009, Biosensors & bioelectronics.

[2]  DoubleTree Hotel Atlanta DEPARTMENT OF HEALTH AND HUMAN SERVICES PUBLIC HEALTH SERVICE CENTERS FOR DISEASE CONTROL AND PREVENTION NATIONAL CENTER FOR INJURY PREVENTION AND CONTROL ADVISORY COMMITTEE FOR INJURY PREVENTION AND CONTROL , 2006 .

[3]  N. Pourmand,et al.  Label-Free Impedance Biosensors: Opportunities and Challenges. , 2007, Electroanalysis.

[4]  B. Nørrung,et al.  Microbial safety of meat in the European Union. , 2008, Meat science.

[5]  J. Harris,et al.  Assembly of Covalently-Coupled Disulfide Multilayers on Gold , 1998 .

[6]  E. Alocilja,et al.  A microfabricated biosensor for detecting foodborne bioterrorism agents , 2005, IEEE Sensors Journal.

[7]  J. Butler Solid supports in enzyme-linked immunosorbent assay and other solid-phase immunoassays. , 2000, Methods.

[8]  Yanbin Li,et al.  Immunobiosensor chips for detection of Escherichia coil O157:H7 using electrochemical impedance spectroscopy. , 2002, Analytical chemistry.

[9]  T. Mexia,et al.  Author ' s personal copy , 2009 .

[10]  J. P. Davis,et al.  A massive outbreak in Milwaukee of cryptosporidium infection transmitted through the public water supply. , 1994, The New England journal of medicine.

[11]  E. Baldrich,et al.  Immunofunctionalisation of gold transducers for bacterial detection by physisorption , 2008, Analytical and bioanalytical chemistry.

[12]  Anthony Turner,et al.  Principles of Bacterial Detection: Biosensors, Recognition Receptors and Microsystems. , 2008 .

[13]  G. Kim,et al.  Nano-particle enhanced impedimetric biosensor for detedtion of foodborne pathogens , 2007 .

[14]  R. Tauxe Emerging foodborne pathogens. , 2002, International journal of food microbiology.

[15]  E. Alocilja,et al.  A high density microelectrode array biosensor for detection of E. coli O157:H7. , 2005, Biosensors & bioelectronics.

[16]  Francesc Xavier Muñoz,et al.  Detection of Escherichia coli and Salmonella typhimurium using interdigitated microelectrode capacitive immunosensors: the importance of transducer geometry. , 2008, Analytical chemistry.

[17]  Dong-Joo Kim,et al.  Phage immobilized magnetoelastic sensor for the detection of Salmonella typhimurium. , 2007, Journal of microbiological methods.

[18]  F Lisdat,et al.  Impedance spectroscopy and biosensing. , 2008, Advances in biochemical engineering/biotechnology.

[19]  Self-assembled monolayers as a base for immunofunctionalisation: unequal performance for protein and bacteria detection , 2008, Analytical and bioanalytical chemistry.

[20]  N. Van Mau,et al.  Protein structural changes induced by their uptake at interfaces. , 2002, Biochimica et biophysica acta.

[21]  Int J Food Microbiol , 2011 .

[22]  R. Jayavel,et al.  Growth and characterization of semiorganic nonlinear optical tetrakis thiourea nickel chloride single crystals , 2007 .

[23]  M. Prodromidis,et al.  Development of an impedimetric immunosensor based on electropolymerized polytyramine films for the direct detection of Salmonella typhimurium in pure cultures of type strains and inoculated real samples. , 2008, Analytica chimica acta.

[24]  Yanbin Li,et al.  Interdigitated array microelectrodes based impedance biosensors for detection of bacterial cells. , 2009, Biosensors & bioelectronics.

[25]  Shankar Balasubramanian,et al.  Lytic phage as a specific and selective probe for detection of Staphylococcus aureus--A surface plasmon resonance spectroscopic study. , 2007, Biosensors & bioelectronics.

[26]  W. Levine,et al.  U.S. Centers for Disease Control and Prevention Guidelines for the Treatment of Sexually Transmitted Diseases: An Opportunity To Unify Clinical and Public Health Practice , 2002, Annals of Internal Medicine.

[27]  Yanbin Li,et al.  A label-free, microfluidics and interdigitated array microelectrode-based impedance biosensor in combination with nanoparticles immunoseparation for detection of Escherichia coli O157:H7 in food samples , 2007 .

[28]  Rosemonde Mandeville,et al.  Bacteriophage-modified microarrays for the direct impedimetric detection of bacteria. , 2008, Analytical chemistry.