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Jump to: navigation, search A biosensor is an analytical device for the detection of an analyte that combines a biological component with a physicochemical detector component. It consists of 3 parts now:
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the sensitive biological element (biological material (e.g. tissue, microorganisms, organelles, cell receptors, enzymes, antibodies, nucleic acids, etc.), a biologically derived material or biomimic component that interacts (binds or recognises) the analyte under study. The biologically sensitive elements can also be created by biological engineering. the transducer or the detector element (works in a physicochemical way; optical, piezoelectric, electrochemical, etc.) that transforms the signal resulting from the interaction of the analyte with the biological element into another signal (i.e., transducers) that can be more easily measured and quantified; biosensor reader device with the associated electronics or signal processors that are primarily responsible for the display of the results in a user-friendly way.[1] This sometimes accounts for the most expensive part of the sensor device, however it is possible to generate a user friendly display that includes transducer and sensitive element(see Holographic Sensor). The readers are usually custom-designed and manufactured to suit the different working principles of biosensors. Known manufacturers of biosensor electronic readers include PalmSens, Gwent Biotechnology Systems and Rapid Labs.

A common example of a commercial biosensor is the blood glucose biosensor, which uses the enzyme glucose oxidase to break blood glucose down. In doing so it first oxidizes glucose and uses two electrons to reduce the FAD (a component of the enzyme) to FADH2. This in turn is oxidized by the electrode (accepting two electrons from the electrode) in a number of steps. The resulting current is a measure of the concentration of glucose. In this case, the electrode is the transducer and the enzyme is the biologically active component. Recently, arrays of many different detector molecules have been applied in so called electronic nose devices, where the pattern of response from the detectors is used to fingerprint a substance.[citation needed]. In the Wasp Hound odor-detector, the mechanical element is a video camera and the biological element is five parasitic wasps who have been conditioned to swarm in response to the presence of a specific chemical.[2] Current commercial electronic noses, however, do not use biological elements. A canary in a cage, as used by miners to warn of gas, could be considered a biosensor. Many of today's biosensor applications are similar, in that they use organisms which respond to toxic substances at a much lower concentrations than humans can detect to warn of the presence of the toxin. Such devices can be used in environmental monitoring, trace gas detection and in water treatment facilities.

Contents

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1 Principles of Detection o 1.1 Photometric o 1.2 Electrochemical o 1.3 Ion Channel Switch o 1.4 Others 2 Applications o 2.1 Glucose monitoring o 2.2 Interferometric Reflectance Imaging Sensor 3 Biosensors in food analysis 4 Surface Attachment of the biological elements 5 See also 6 References 7 External links

[edit] Principles of Detection

[edit] Photometric
Many optical biosensors based on the phenomenon of surface plasmon resonance (SPR) are evanescent wave techniques. This utilises a property of gold and other materials; specifically that a thin layer of gold on a high refractive index glass surface can absorb laser light, producing electron waves (surface plasmons) on the gold surface. This occurs only at a specific angle and wavelength of incident light and is highly dependent on the surface of the gold, such that binding of a target analyte to a receptor on the gold surface produces a measurable signal. Surface plasmon resonance sensors operate using a sensor chip consisting of a plastic cassette supporting a glass plate, one side of which is coated with a microscopic layer of gold. This side contacts the optical detection apparatus of the instrument. The opposite side is then contacted with a microfluidic flow system. The contact with the flow system creates channels across which reagents can be passed in solution. This side of the glass sensor chip can be modified in a number of ways, to allow easy attachment of molecules of interest. Normally it is coated in carboxymethyl dextran or similar compound. Light of a fixed wavelength is reflected off the gold side of the chip at the angle of total internal reflection, and detected inside the instrument. This induces the evanescent wave to penetrate through the glass plate and some distance into the liquid flowing over the surface. The refractive index at the flow side of the chip surface has a direct influence on the behaviour of the light reflected off the gold side. Binding to the flow side of the chip has an effect on the refractive index and in this way biological interactions can be measured to a high degree of sensitivity with some sort of energy. The refractive index of the medium near the surface

changes when biomolecules attach to the surface, and the SPR angle varies as a function of this change. Other evanescent wave biosensors have been commercialised using waveguides where the propagation constant through the waveguide is changed by the absorption of molecules to the waveguide surface. One such example, Dual Polarisation Interferometry uses a buried waveguide as a reference against which the change in propagation constant is measured. Other configurations such as the Mach-Zehnder have reference arms lithographically defined on a substrate. Higher levels of integration can be achieved using resonator geometries where the resonant frequency of a ring resonator changes when molecules are absorbed. Other optical biosensors are mainly based on changes in absorbance or fluorescence of an appropriate indicator compound and do not need a total internal reflection geometry. For example, a fully operational prototype device detecting casein in milk has been fabricated. The device is based on detecting changes in absorption of a gold layer.[3] A widely used research tool, the micro-array, can also be considered a biosensor. Nanobiosensors use a immobilized bioreceptor probe that is selective for target analyte molecules. Nanomaterials are exquisitely sensitive chemical and biological sensors. Nanoscale materials demonstrate unique properties. Their large surface area to volume ratio can achieve rapid and low cost reactions, using a variety of designs.[4] Biological biosensors often incorporate a genetically modified form of a native protein or enzyme. The protein is configured to detect a specific analyte and the ensuing signal is read by a detection instrument such as a fluorometer or luminometer. An example of a recently developed biosensor is one for detecting cytosolic concentration of the analyte cAMP (cyclic adenosine monophosphate), a second messenger involved in cellular signaling triggered by ligands interacting with receptors on the cell membrane.[5] Similar systems have been created to study cellular responses to native ligands or xenobiotics (toxins or small molecule inhibitors). Such "assays" are commonly used in drug discovery development by pharmaceutical and biotechnology companies. Most cAMP assays in current use require lysis of the cells prior to measurement of cAMP. A live-cell biosensor for cAMP can be used in non-lysed cells with the additional advantage of multiple reads to study the kinetics of receptor response.

[edit] Electrochemical
Electrochemical biosensors are normally based on enzymatic catalysis of a reaction that produces or consumes electrons (such enzymes are rightly called redox enzymes). The sensor substrate usually contains three electrodes; a reference electrode, a working electrode and a counter electrode. The target analyte is involved in the reaction that takes place on the active electrode surface, and the reaction may cause either electron transfer across the double layer (producing a current) or can contribute to the double layer potential (producing a voltage). We can either measure the current (rate of flow of electrons is now proportional to the analyte concentration) at a fixed potential or the potential can be measured at zero current (this gives a logarithmic response). Note that potential of the working or active electrode is space charge sensitive and this is often used. Further, the label-free and direct electrical detection of small peptides and proteins

is possible by their intrinsic charges using biofunctionalized ion-sensitive field-effect transistors.[6] Another example, the potentiometric biosensor, (potential produced at zero current) gives a logarithmic response with a high dynamic range. Such biosensors are usually often made by screen printing the electrode patterns on a plastic substrate, coated with a conducting polymer and then some protein (enzyme or antibody) is attached. They have only two electrodes and are extremely sensitive and robust. They enable the detection of analytes at levels previously only achievable by HPLC and LC/MS and without rigorous sample preparation. All biosensors usually involve minimal sample preparation as the biological sensing component is highly selective for the analyte concerned. The signal is produced by electrochemical and physical changes in the conducting polymer layer due to changes occurring at the surface of the sensor. Such changes can be attributed to ionic strength, pH, hydration and redox reactions, the latter due to the enzyme label turning over a substrate ([2]). Field effect transistors, in which the gate region has been modified with an enzyme or antibody, can also detect very low concentrations of various analytes as the binding of the analyte to the gate region of the FET cause a change in the drain-source current.

[edit] Ion Channel Switch

ICS - channel open

ICS - channel closed The use of ion channels has been shown to offer highly sensitive detection of target biological molecules.[7] By imbedding the ion channels in supported or tethered bilayer membranes (tBLM) attached to a gold electrode, an electrical circuit is created. Capture molecules such as antibodies can be bound to the ion channel so that the binding of the target molecule controls the ion flow through the channel. This results in a measurable change in the electrical conduction which is proportional to the concentration of the target. An Ion Channel Switch (ICS) biosensor can be created using gramicidin, a dimeric peptide channel, in a tethered bilayer membrane.[8] One peptide of gramicidin, with attached antibody, is mobile and one is fixed. Breaking the dimer stops the ionic current through the membrane. The magnitude of the change in electrical signal is greatly increased by separating the membrane from the metal surface using a hydrophilic spacer.

Quantitative detection of an extensive class of target species, including proteins, bacteria, drug and toxins has been demonstrated using different membrane and capture configurations.[9][10]

[edit] Others
Piezoelectric sensors utilise crystals which undergo an elastic deformation when an electrical potential is applied to them. An alternating potential (A.C.) produces a standing wave in the crystal at a characteristic frequency. This frequency is highly dependent on the elastic properties of the crystal, such that if a crystal is coated with a biological recognition element the binding of a (large) target analyte to a receptor will produce a change in the resonance frequency, which gives a binding signal. In a mode that uses surface acoustic waves (SAW), the sensitivity is greatly increased. This is a specialised application of the Quartz crystal microbalance as a biosensor. Thermometric and magnetic based biosensors are rare.

[edit] Applications
There are many potential applications of biosensors of various types. The main requirements for a biosensor approach to be valuable in terms of research and commercial applications are the identification of a target molecule, availability of a suitable biological recognition element, and the potential for disposable portable detection systems to be preferred to sensitive laboratorybased techniques in some situations. Some examples are given below:
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Glucose monitoring in diabetes patients historical market driver Other medical health related targets Environmental applications e.g. the detection of pesticides and river water contaminants such as heavy metal ions[11] Remote sensing of airborne bacteria e.g. in counter-bioterrorist activities Detection of pathogens[12] Determining levels of toxic substances before and after bioremediation Detection and determining of organophosphate Routine analytical measurement of folic acid, biotin, vitamin B12 and pantothenic acid as an alternative to microbiological assay Determination of drug residues in food, such as antibiotics and growth promoters, particularly meat and honey. Drug discovery and evaluation of biological activity of new compounds. Protein engineering in biosensors [13] Detection of toxic metabolites such as mycotoxins [14]

[edit] Glucose monitoring

Main article: Blood glucose monitoring Commercially available gluocose monitors rely on amperometric sensing of glucose by means of glucose oxidase, which oxidises glucose producing hydrogen peroxide which is detected by the

electrode. To overcome the limitation of ameperometric sensors, a flurry of research is present into novel sensing methods, such as fluorescent glucose biosensors.

[edit] Interferometric Reflectance Imaging Sensor

The Interferometric Reflectance Imaging Sensor (IRIS) was developed by the Unlu research group at Boston University for the purpose of label-free biosensing. Using simple lenses and low-powered, coherent LEDs, the device offers exquisite sensitivity and reproducibility and is able to image with remarkable resolution beyond the classical diffraction limit. This relatively cheap solution also presents minimal hazards when compared to a laser illumination source. One specific form of photometric biosensing technique developed by researchers at Boston University is interferometric reflectance imaging. Using optical interference techniques, imaging of antibodies were successfully performed. This was achieved without altering the antibody structure or using bio-markers such as fluorescent proteins. The basis of this technique stems solely from optical interference. By using a reflective substrate such as silicon, light reflected from proteins will interfere with light reflected from the substrate. In result, interference patterns are generated that alter the intensity of the reflected light. This phenomena is measurable by a camera.

Proteins have indices of refraction based on their concentration. When light is shined on the proteins, a portion of the light is transmitted through the molecules and reflected off the silicon's surface. The interference of the light initially reflected off the proteins and the light reflected off the surface of the silicon will have a relative phase difference (after being transmitted back through the protein) contributing to a wavelength-dependent sinusoidal variation in the total amount of reflected light (captured by the imaging device). The Interferometric Reflectance Imaging Sensor (IRIS) was developed by the Unlu research group at Boston University for the purpose of label-free biosensing. Using simple lenses and low-powered, coherent LEDs, the device offers exquisite sensitivity and reproducibility and is able to image with remarkable resolution beyond the classical diffraction limit. This relatively cheap solution also presents minimal hazards when compared to a laser illumination source. The IRIS operates solely on optical reflection. The ability for it to image with extremely high spatial resolution stems from the integration of a diffuser into the design of the microscope. The diffuser randomizes the directionality of the light from a single LED source (called Khler illumination) which allows for sharp focusing of incident light without back-imaging the source in the image projection. Practical uses of this device include the detection of bacterial and viral infections in underdeveloped countries. When pathogen specific growth factors are introduced into a microarray, only spots with the targeted pathogens will grow and increase in concentration. In turn, this dictates a change in the reflected intensity compared to pre-growth. Thus, by measuring

how reflectance changes over time, unknown pathogens and their growth rates can be easily characterized and identified.

[edit] Biosensors in food analysis

There are several applications of biosensors in food analysis. In food industry optic coated with antibodies are commonly used to detect pathogens and food toxins. The light system in these biosensors has been fluorescence, since this type of optical measurement can greatly amplify the signal. A range of immuno- and ligand-binding assays for the detection and measurement of small molecules such as water-soluble vitamins and chemical contaminants (drug residues) such as sulfonamides and Beta-agonists have been developed for use on SPR based sensor systems, often adapted from existing ELISA or other immunological assay. These are in widespread use across the food industry.

[edit] Surface Attachment of the biological elements

An important part in a biosensor is to attach the biological elements (small molecules/protein/cells) to the surface of the sensor (be it metal, polymer or glass). The simplest way is to functionalize the surface in order to coat it with the biological elements. This can be done by polylysine, aminosilane, epoxysilane or nitrocellulose in the case of silicon chips/silica glass. Subsequently the bound biological agent may be for example fixed by Layer by layer depositation of alternatively charged polymer coatings[15] Alternatively three dimensional lattices (hydrogel/xerogel) can be used to chemically or physically entrap these (where by chemically entraped it is meant that the biological element is kept in place by a strong bond, while physically they are kept in place being unable to pass through the pores of the gel matrix). The most commonly used hydrogel is sol-gel, a glassy silica generated by polymerization of silicate monomers (added as tetra alkyl orthosilicates, such as TMOS or TEOS) in the presence of the biological elements (along with other stabilizing polymers, such as PEG) in the case of physical entrapment.[16] Another group of hydrogels, which set under conditions suitable for cells or protein, are acrylate hydrogel, which polymerize upon radical initiation. One type of radical initiator is a peroxide radical, typically generated by combining a persulfate with TEMED (Polyacrylamide gel are also commonly commonly used for protein electrophoresis),[17] alternatively light can be used in combination with a photoinitiator, such as DMPA (2,2-dimethoxy-2-phenylacetophenone).[18] Smart materials that mimic the biological components of a sensor can also be classified as biosensors using only the active or catalytic site or analogous configurations of a biomolecule.[19]

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Conducting Polymers and Their Applications to Biosensors: Emphasizing on

Foodborne Pathogen Detection

Khalil Arshak, Vijayalakshmi Velusamy, Olga Korostynska, Kamila Oliwa-Stasiak, and Catherine Adley
AbstractDetection of microbial pathogens in food is the solution to the prevention and recognition of problems related to health and safety. New biomolecular approaches for foodborne pathogen detection are being developed to improve the biosensor characteristics such as sensitivity and selectivity, also which is rapid, reliable, cost-effective, and suitable for in situ analysis. Recently, conducting polymers have drawn attention in the development of biosensors. The electrically conducting polymers have numerous features, which allow them to act as excellent materials for immobilization of biomolecules. Also, their unique properties make them appealing alternatives for specific materials currently employed for the fabrication of biosensors. Therefore, this paper presents a comprehensive literature review detailing the salient features of conducting polymers and their application to biosensors with an emphasis on foodborne pathogen detection. Index TermsBiosensors, conducting polymers, foodborne pathogen detection, identification, immobilization, quantification, transducer.

I. INTRODUCTION

LTHOUGH food safety has dramatically improved

overall, progress is uneven and foodborne outbreaks from microbial contamination, chemicals and toxins are common in many countries [1]. Therefore, the role of pathogen detection technology is vital, which is the key to the prevention and identification of problems related to health and safety. But still, a detection technique which is reliable, rapid, accurate, simple, sensitive, selective, cost effective and also suitable for in situ analysis is yet to be developed. Conducting polymers have unique properties which make them appealing alternatives for specific materials currently employed for the fabrication of biosensors. Since, the literature related to application of conducting polymers to biosensors is vast; this paper reports mainly on the detection, identification and quantification of foodborne pathogens.
Manuscript received October 21, 2008; revised April 28, 2009; accepted August 31, 2009. Current version published November 04, 2009. The associate editor coordinating the review of this paper and approving it for publication was Prof. Massood Atashbar. This work was supported in part by the Science Foundation Ireland (SFI) Research Frontiers Program under 07RPF-ENEF500. K. Arshak, V. Velusamy, and O. Korostynska are with the Department of Electronic and Computer Engineering, University of Limerick, Limerick, Ireland (e-mail: khalil.arshak@ul.ie; viji.subha@ul.ie; olga.korostynska@ul.ie). K. Oliwa-Stasiak and C. Adley are with the Microbiology Laboratory, Department of Chemical and Environmental Sciences, University of Limerick, Limerick, Ireland (e-mail: kamila.oliwa@ul.ie; Catherine.adley@ul.ie). Digital Object Identifier 10.1109/JSEN.2009.2032052 Fig. 1. Polyacetylene chain.

A. What Makes Polymers Conductive? Polymers (plastics) are known to have good insulating properties and are among the most used materials in the modern world. However, it has been discovered that there are some polymers which have conducting properties. Conducting polymers are polymer materials with metallic and semiconductor characteristics, a combination of properties not exhibited by any other known material. A key property of a conductive polymer is the

presence of conjugated double bonds along the backbone of the polymer. In conjugation, the bonds between the carbon atoms are alternately single and double. For example, structure of polyacetylene is shown in Fig. 1. Since the electrons in a conjugated system are only loosely bound, electron flow may be possible. Every bond contains a localized sigma bond which forms a strong chemical bond. In addition, every double bond also contains a less strongly localized pi bond which is weaker. These enable the electrons to be delocalized over the whole system and so be shared by many atoms. This means that the delocalized electrons may move around the whole system. However, conjugation is not enough to make the polymer material conductive. In addition, the polymer material needs to be doped for electron flow to occur. Doping is either the addition of electrons (reduction reaction) or the removal of electrons (oxidation reaction) from the polymer. An oxidation doping (removal of electrons) can be done using iodine. The iodine attracts an electron from the polymer from one of the bonds. Once doping has occurred, the electrons in the -bonds are able to jump around the polymer chain. As the electrons are moving along the molecule, electric current occurs. For better conductivity the molecules must be well ordered and closely packed to limit the distance jumped by the electrons. The conductivity of conducting polymers can be tuned by chemical manipulation of the polymer backbone, by the nature of the dopant, by the degree of doping, and by blending with other polymers [2].
. .

Introduction to Biosensors A biosensor is an analytical device, which converts a biological response into an electrical signal. It consists of two main components: a bioreceptor or biorecognition element, which recognizes the target analyte and a transducer, for converting the recognition event into a measurable electrical signal. A bioreceptor can be a tissue, microorganisms, organelles, cells, enzymes, antibodies, nucleic acids and biomimic and the transduction may be optical, electrochemical, thermometric, piezoelectric, magnetic and micromechanical or combinations of one or more of the above techniques. Fig. 4 shows schematic diagram of a biosensor. The bioreceptor recognizes the target analyte and the corresponding biological responses are then converted into equivalent electrical signals by the transducer. The amplifier in the biosensor responds to the small input signal from the transducer and delivers a large output signal that contains the essential waveform features of an input signal. The amplified signal is then processed by the signal processor where it can later be stored, displayed and analyzed. Biosensors have been widely applied to a variety of analytical problems in medicine, the environment, food, process industries, security, and defense.

SIGNIFICANCE OF CONDUCTING POLYMERS TO BIOSENSORS Conducting polymers have been used as a transducer in biological sensors due to its attractive properties and their use in

biosensors has grown over the past decade. Some of the attractive features of conducting polymers for biosensor applications include: Availability of varied range of monomer types. Availability synthetic analogues of monomers. Composites can be prepared combing conducting polymers with nonconducting polymers or with nonpolymer materials such carbon, carbon nanotubes, metals, etc. It can be prepared both electrochemically and chemically. It can be prepared in a range of soluble and insoluble forms. It has unique electrical, electronic, magnetic and optical properties. Compliance with micro and nanoscale fabrication. Compatibility with diverse range of fabrication techniques such as electrochemical, optical, mass-based, etc. Biomaterials such as enzymes, antibodies, whole cells, and nucleic acids can be incorporated into the polymer matrix. Strong biomolecular interactions. Low detection limits. Enhanced sensitivity (when used as a composite material with nanoparticles). Reversible responses at ambient temperatures. Cost effectiveness.

Classifications of Biosensor Biosensors can be classified by their bioreceptor or their transducer type. Fig. 6 shows the biosensor classifications. Bioreceptors: Classified into five different major categories.

These categories include antibody/antigen, enzymes, nucleic acids/DNA/RNA, cellular structures/cells, and biomimetic. The enzymes, antibodies, and nucleic acids are the main classes of bioreceptors which are widely used in biosensor applications. Though the enzymes are one of the biorecognition elements, they are mostly used to function as labels than the actual bioreceptor. Transducers: The transducer plays an important role in the detection process of a biosensor. In case of conducting polymerbased biosensor, the conductive polymer acts as a transducer that converts the biological signal to an electrical signal. Biosensors can also be classified based upon the transduction methods they employ.Wide varieties of transduction methods have been developed in the past decade for the detection of foodborne pathogens. Although there are new types of transducers constantly being developed for use in biosensors, the transduction methods such as optical, electrochemical, and mass based are given importance here since these are the most popular and common methods. Each of these three main classes contains many different subclasses and they can be further divided into label and label-free (nonlabeled) methods, where, the labeled methods depend on the detection of a specific label (e.g., fluorescence) and the label-free detection is based on the direct measurement of a product developing during the biochemical reactions on a transducer surface.

IX. CONCLUSION Although many conducting polymer-based biosensors for foodborne pathogen detection has been published in recent years, only very few applications to food samples have been reported to date. Among other conducting polymers, polyaniline, and polypyrrole were most extensive reported and polypyrrole was found to be a better choice for DNA biosensors. Foodborne pathogen detection based on electrochemical detection is preferred more than optical detection. To fabricate a conducting polymer-based biosensor, it is not only important to consider the properties of conducting polymers, but the method of
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1950 IEEE SENSORS JOURNAL, VOL. 9, NO. 12, DECEMBER 2009

immobilization plays a vital role. Therefore, by making use of conducting polymers with suitable immobilization technique, it is possible to develop a novel biosensor to detect foodborne pathogen detection in real time. Also, the use of nanomaterials/ nanoparticles as a composite with conducting polymer would lead to the fabrication of biosensors and many other nanoelectronic devices and the basic requirements of biosensor like, enhanced sensitivity and very low detection limit can also achieved.

VI. CONDUCTING POLYMER BASED BIOSENSOR FOR FOODBORNE PATHOGEN DETECTION A. Electrochemical Biosensor Electrochemical-based detection methods are another possible

means of transduction that has been used for identification and quantification of foodborne pathogens. Electrochemical biosensors can be classified into amperometric, potentiometric, impedimetric and conductometric, based on the observed parameters such as current, potential, impedance, and conductance, respectively. Although the electrochemical detection has several advantages like low cost, ability to work with turbid samples, and easy miniaturization, their sensitivity and selectivity is slightly limited when compared to optical detection. However, electrochemical biosensors were found coupled with other biosensing techniques for enhanced detection. Also, electrochemical polymerization is an effective technique for the deposition of conducting polymers onto various substrates and following are the advantages of electrochemical deposition. i) It allows the reproducible and precise formation of a conducting polymer coating over surfaces whatever their size and geometry. ii) Growth of the conducting polymer can be controlled. iii) The polymeric films are stable in organic and aqueous solvents. Amperometric Detection: Amperometric transduction is the most common electrochemical detection method which has been used for pathogen detection and it has superior sensitivity than potentiometic method. In amperometric-based detection, the sensor potential is set at a value where the analyte produces current. Thus, the applied potential serves as the driving force for the electron transfer reaction, and the current produced is a direct measure of the rate of electron transfer. Minett et al. [34], coupled conducting polymers and the electron mediators for the detection of Listeria monocytogenes. Different signal generation techniques were investigated for the detection of L. monocytogenes at both bare and polymer (polypyrrole)-modified electrodes. The paper reported that conventional amperometry at an antibody-containing polypyrrole film electrode was found to be unsuccessful in detecting levels below cells/mL. More successful was the coupling of a covalently modified film with the use of electron mediators in a single device. It was found that the use of conventional techniques such as voltammetry was successful in detecting high levels of micro-organisms ( cells/mL Listeria). However, these techniques suffered from a lack of reproducibility between polymer films and an inability to detect small viable numbers. Improving the immobilization of the antibody (covalent attachment) met with little success when using voltammetry. Although detection was possible, detecting low levels of micro-organisms was beyond the capabilities of the technique in these experiments. The use of Toluidine Blue at a bare glassy carbon electrode also proved successful in detecting Listeria at concentrations of cells/mL. The method was not specific as responses were also obtained for other micro-organisms, such as E. coli and Salmonella. The
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TYPES OF CONDUCTING POLYMERS Only emeraldine polymer exhibits conductivity. If emeraldinebase polymer is treated with acidic solution (either organic or inorganic protonic acids) with pH lower than 4, it is converted to emeraldine salt form which is the conducting form of the emeraldine polymer. If the polymer is treated with a solution with pH greater than 4, the polymer becomes insulator [5]. Common conducting polymers are poly(acetylene)s, poly (pyrrole)s, poly(thiophene)s, poly(terthiophene)s, poly(aniline) s, poly(fluorine)s, poly(3-alkylthiophene)s, polytetrathiafulvalenes, polynapthalenes, poly( -phenylene sulfide), poly( -phenylenevinylene)s, poly(3,4-ethylenedioxythiophene), polyparaphenylene, polyazulene, polyparaphenylene sulfide, polycarbazole, and polydiaminonaphthalene [6]. Among the various conducting polymers polyaniline, polythiophene, and polypyrrole, are biocompatible and hence, cause minimal and reversible disturbance to the working environment and protect electrodes from fouling [7]. However, only polyaniline, polypyrrole are most extensive used in biosensors for foodborne pathogen detection. Recently, conducting polymer nanocomposites which have enhanced properties has been developed to overcome the inherent limitations of pure conducting polymers [8]. The use of conducting polymer nanocomposites/nanoparticles could greatly improve diffusion since they have much greater exposed surface area and as a result of this the basic characteristics of a biosensor like low detection limit get enhanced. The oriented microstructure and the high surface area also facilitate high biomolecule loading and hence highly sensitive detection is possible. Moreover, the relative stability is increased due to efficient bonding of biomolecule on the transducer surface which improves reproducibility. These materials are especially important because of their bridging role between the world of conducting polymers and that of nanoparticles [9]. The promising features of conducting polymer nanocomposites are also discussed in literature [10][15]. Pal and Alocilja [14] has developed electrically active polyaniline coated magnetic (EAPM) nanoparticle-based biosensor for the detection of Bacillus anthracis endospores in contaminated food samples. The 100 nm-diameter EAPM nanoparticles are synthesized from aniline monomer (made electrically active by acid doping) coating the surface of gamma iron oxide cores. Experimental results indicate that the biosensor is able to detect B. anthracis spores at concentrations as low as spores/ml from the samples with a total detection time of 16 min. Ma et al. [16] used a nanocomposite of poly(anilineboronic acid), a self-doped polyaniline, with ss-DNA-wrapped single-walled carbon nanotubes (ss-DNA/SWNTs) to fabricate on a gold electrode by in situ electrochemical polymerization of 3-aminophenylboronic acid monomers in the presence of ssDNA/SWNTs. The paper reported that the sensitivity increased four orders of magnitude when nanocomposite was used to detect nanomolar concentrations of dopamine compared to the detection at an electrode modified with only poly(anilineboronic acid).

A. Polyaniline

Among the other organic conjugated polymers, polyaniline received considerable attention in recent years due to its high conductivity in the doped state (up to 1 S/cm), the polymerization reaction can be easily controlled to give high yields, the monomer is inexpensive, its excellent environmental and thermal stability as well as the electrochemical stability [5], [17]. The structure of polyaniline is shown in Fig. 2. The emeraldine salt form of polyaniline can be easily obtained by HCl doping which imparts good environmental and thermal stability, while enhancing relatively high conductivity [17]. The responsive nature of polyaniline is highly dependent on processing conditions, film composition, and morphology. The mechanisms employed in producing polyaniline films for chemical/biological sensing determine the level of electrical conductivity, overall structure, and stability [18]. Langer and
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1944 IEEE SENSORS JOURNAL, VOL. 9, NO. 12, DECEMBER 2009

Fig. 3. Structure of polypyrrole.

Langer developed a nanobiodetector to observe the specific changes in the electrical conductivity of a polyaniline micro and nanofibril network in the presence of micro-organisms [19]. It has been observed that the electrical conductivity of polyaniline micro and nanofibrils is strongly dependent on the temperature and the chemical environment. The paper reported that the electrical response depends on the number of cells deposited on the polyaniline nanonetwork, and is specific to different kinds of micro-organisms with very low detection limit. Among other conducting polymers, polyaniline is often used as an immobilizing substrate for biomolecules [7] and it has proven particularly useful in the development of biosensors [20]. However, the intractability of polyaniline has limited its utilization in commercial biosensors, especially in its pure, inherently conductive form [21], [22]. Dispersion of polyaniline is one of the interesting ways to improve the processability of Polyaniline. Morrin et al. [21] developed a simple fabrication method for a biosensor by depositing polyaniline nanoparticles, where the dispersions were cast onto disposable screen-printed electrodes by means of drop-coating. The paper reported that the method was shown to be an effective biosensor format. Also, the signal-to-background (S/B) was comparable to the biosensor developed using electrodeposited nanoparticles [20].

B. Polypyyrole
Polypyrrole is one of the most attractive conducting polymers with some special electrical properties. These properties originate from the fact that Polypyrrole is an intrinsically conducting polymer and can be synthesized to have conductivities up to 1000 S/cm which approaches the conductivity of metals. Electrical conduction in polypyrrole is the result of electron movement within delocalized orbitals and positive charge defects known as polarons [23]. In conjugated polymers other than polyacetylene, electrons added or removed from the delocalized -bonded backbone initially produce polarons (radical ions coupled to a spatially extended distortion of the bond lengths) which subsequently combine to form dianions or dications (spinless bipolarons), respectively [24]. In polyacetylene, anions and cations are produced during charge transfer. Most

practical types of Polypyrrole have conductivities in the range of 1100 S/cm [9]. The structure of polypyrrole is shown in Fig. 3. Polypyrrole is used mostly in biosensors and immunosensors because of the best biocompatibility and the ease of immobilization of various biologically active compounds [25]. To detect bio-analytes at a physiological pH, biosensing materials must be electroactive in neutral environments, unlike polyaniline and polythiophene. To overcome this problem, polypyrrole is found to be very attractive because it can be more easily deposited from neutral pH aqueous solutions of pyrrolemonomers [26].
Fig. 4. Schematic diagram of a biosensor.

DNA can form a strong bond with polypyrrole based on the interchanging of dopant DNA molecules [27] and hence it has attracted attention of various researchers for application of DNA biosensors [28]. Recently, Zanuy and Aleman [29] examined the ability of pyrrole and thiophene, which are the monomeric units of polypyrrole and polythiophene, respectively, to interact with the methylated analogues of DNA bases, 9-methyladenine (mA), 9-methylguanine (mG), 1-methylcytosine (mC), and 1-methylthymine (mT) through specific hydrogen-bonding interactions. Results evidenced that pyrrole is a strong proton donor, able to form very stable complexes with mA, mG, mC, and mT. Moreover, the specificity of pyrrole to methylated nucleic acids is very remarkable. On the other hand, the sulfur of thiophene is a very weak proton acceptor as was revealed by the fact that no hydrogen-bonded complex was formed with mA, mC, and mT. The paper reported that the results were fully consistent with the high affinity of polypyrrole toward plasmid DNA, as well as with the ability of this polymer to form specific interactions with well-defined nucleotide sequences protecting DNA from enzymatic digestion.

Schematic diagram of a biosensor.