Label-free impedimetric biosensor for Salmonella Typhimurium detection based on poly [pyrrole-co-3-carboxyl-pyrrole] copolymer supported aptamer.

The Gram-negative bacterium, Salmonella Typhimurium (S. Typhimurium) is a food borne pathogen responsible for numerous hospitalisations and deaths all over the world. Conventional detection methods for pathogens are time consuming and labour-intensive. Hence, there is considerable interest in faster and simpler detection methods. Polypyrrole-based polymers, due to their intrinsic chemical and electrical properties, have been demonstrated to be valuable candidates for the fabrication of chemo/biosensors and functional surfaces. Similarly aptamers have been shown to be good alternatives to antibodies in the development of affinity biosensors. In this study, we report on the combination of poly [pyrrole-co-3-carboxyl-pyrrole] copolymer and aptamer for the development of a label-less electrochemical biosensor suitable for the detection of S. Typhimurium. Impedimetric measurements were facilitated by the effect of the aptamer/target interaction on the intrinsic conjugation of the poly [pyrrole-co-3-carboxyl-pyrrole] copolymer and subsequently on its electrical properties. The aptasensor detected S. Typhimurium in the concentration range 10(2)-10(8) CFU mL(-1) with high selectivity over other model pathogens and with a limit of quantification (LOQ) of 100 CFU mL(-1) and a limit of detection (LOD) of 3 CFU mL(-1). The suitability of the aptasensor for real sample detection was demonstrated via recovery studies performed in spiked apple juice samples. We envisage this to be a viable approach for the inexpensive and rapid detection of pathogens in food, and possibly in other environmental samples.

[1]  Ronghui Wang,et al.  Rapid detection of Escherichia coli O157:H7 and Salmonella Typhimurium in foods using an electrochemical immunosensor based on screen-printed interdigitated microelectrode and immunomagnetic separation. , 2016, Talanta.

[2]  Wilfred Chen,et al.  Reversible conversion of conducting polymer films from superhydrophobic to superhydrophilic. , 2005, Angewandte Chemie.

[3]  A. Turner,et al.  Tunable conjugated polymers for bacterial differentiation , 2015 .

[4]  Jiri Janata,et al.  Label-free DNA hybridization probe based on a conducting polymer. , 2003, Journal of the American Chemical Society.

[5]  H. Bagheri,et al.  Conductive polymer-based microextraction methods: a review. , 2013, Analytica chimica acta.

[6]  Christian Soeller,et al.  Label-free electrochemical DNA sensor based on functionalised conducting copolymer. , 2005, Biosensors & bioelectronics.

[7]  A. Abbaspour,et al.  Aptamer-conjugated silver nanoparticles for electrochemical dual-aptamer-based sandwich detection of staphylococcus aureus. , 2015, Biosensors & bioelectronics.

[8]  H. Alakomi,et al.  Salmonella importance and current status of detection and surveillance methods , 2009 .

[9]  Christine E. Schmidt,et al.  Conducting polymers in biomedical engineering , 2007 .

[10]  Yi Shi,et al.  A nanostructured conductive hydrogels-based biosensor platform for human metabolite detection. , 2015, Nano letters.

[11]  Takaomi Kobayashi,et al.  A novel highly sensitive humidity sensor based on poly(pyrrole-co-formyl pyrrole) copolymer film: AC and DC impedance analysis , 2015 .

[12]  Christian Soeller,et al.  Label-free detection of DNA hybridization based on a novel functionalized conducting polymer. , 2007, Biosensors & bioelectronics.

[13]  A. Yassar,et al.  Electrochemical probing of DNA based on oligonucleotide-functionalized polypyrrole. , 2001, Biomacromolecules.

[14]  Edwin W H Jager,et al.  Mechanical stimulation of epithelial cells using polypyrrole microactuators. , 2011, Lab on a chip.

[15]  Yuzuru Takamura,et al.  Labelless impedance immunosensor based on polypyrrole-pyrolecarboxylic acid copolymer for hCG detection. , 2011, Talanta.

[16]  Giyoung Kim,et al.  A microfluidic nano-biosensor for the detection of pathogenic Salmonella. , 2015, Biosensors & bioelectronics.

[17]  C. Soeller,et al.  Conducting polymers for electrochemical DNA sensing. , 2009, Biomaterials.

[18]  David E. Williams,et al.  High-sensitivity, label-free DNA sensors using electrochemically active conducting polymers. , 2011, Analytical chemistry.

[19]  C. O’Sullivan,et al.  Cystic fibrosis: a label-free detection approach based on thermally modulated electrochemical impedance spectroscopy , 2010, Analytical and bioanalytical chemistry.

[20]  S. Cartmell,et al.  Conductive polymers: towards a smart biomaterial for tissue engineering. , 2014, Acta biomaterialia.

[21]  Inamuddin,et al.  A conducting polymer/ferritin anode for biofuel cell applications , 2009 .

[22]  Faridah Salam,et al.  Detection of Salmonella typhimurium using an electrochemical immunosensor. , 2009, Biosensors & bioelectronics.

[23]  G. S. Zamay,et al.  Aptamer-based viability impedimetric sensor for bacteria. , 2012, Analytical chemistry.

[24]  Lee-Ann Jaykus,et al.  Selection, characterization, and application of DNA aptamers for the capture and detection of Salmonella enterica serovars. , 2009, Molecular and cellular probes.

[25]  Edwin Jager,et al.  Controlling the electro-mechanical performance of polypyrrole through 3- and 3,4-methyl substituted copolymers , 2015 .

[26]  Claude Martelet,et al.  Electrochemical impedance probing of DNA hybridisation on oligonucleotide-functionalised polypyrrole. , 2005, Talanta.

[27]  E. Smela,et al.  Microfabricating conjugated polymer actuators. , 2000, Science.

[28]  Wei Chen,et al.  Sensitive human interleukin 5 impedimetric sensor based on polypyrrole-pyrrolepropylic acid-gold nanocomposite. , 2008, Analytical chemistry.

[29]  John G. Bruno,et al.  Application of DNA Aptamers and Quantum Dots to Lateral Flow Test Strips for Detection of Foodborne Pathogens with Improved Sensitivity versus Colloidal Gold , 2014, Pathogens.

[30]  Geoffrey M. Spinks,et al.  Conductive Electroactive Polymers: Intelligent Polymer Systems , 2009 .

[31]  Zhouping Wang,et al.  An aptamer-based electrochemical biosensor for the detection of Salmonella. , 2014, Journal of microbiological methods.

[32]  J. Riu,et al.  Immediate detection of living bacteria at ultralow concentrations using a carbon nanotube based potentiometric aptasensor. , 2009, Angewandte Chemie.

[33]  T. S. West Analytical Chemistry , 1969, Nature.

[34]  G. S. Zamay,et al.  Aptamer-based impedimetric sensor for bacterial typing. , 2012, Analytical chemistry.

[35]  Pranjal Chandra,et al.  Label-free detection of kanamycin based on the aptamer-functionalized conducting polymer/gold nanocomposite. , 2012, Biosensors & bioelectronics.

[36]  S. Phanichphant,et al.  Electrochemically Fabricated Pyrrole Copolymer Thin Films and Their Electroactivity in Neutral Aqueous Solution , 2013 .

[37]  J. Riu,et al.  Label-free detection of Staphylococcus aureus in skin using real-time potentiometric biosensors based on carbon nanotubes and aptamers. , 2012, Biosensors & bioelectronics.

[38]  Huangxian Ju,et al.  A Rapid and Sensitive Aptamer-Based Electrochemical Biosensor for Direct Detection of Escherichia Coli O111 , 2012 .

[39]  F. Garnier,et al.  Conjugated polymer-based DNA chip with real time access and femtomol detection threshold , 2007 .