RESUMO
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.
Assuntos
Bactérias/isolamento & purificação , Bacteriófagos , Técnicas Biossensoriais/métodos , Anticorpos Antibacterianos , Carga Bacteriana , Bacteriófago T4 , Espectroscopia Dielétrica/métodos , Escherichia coli K12/imunologia , Escherichia coli K12/isolamento & purificação , Ouro , Lactobacillus plantarum/isolamento & purificação , Microeletrodos , Espectroscopia de Infravermelho com Transformada de Fourier , Microbiologia da ÁguaRESUMO
In recent years, conducting polymers combined with metallic nanoparticles have been paid more attention due to their potential applications in microelectronics, microsystems, optical sensors and photoelectronic chemistry. The work presented in this paper describes the preparation and characterization of a nanocomposite composed by a thin polypyrrole (PPy) film covered with an assembly of magnetic nanoparticles (NPs). The magnetic particles were immobilized on PPy films under appropriate magnetic field in order to control their organization on the PPy film and finally to improve the sensitivity of the system in potential sensing applications. The electrical properties and morphology of the resulting PPy film and the PPy film/NPs composite were characterized with cyclic voltammetry, impedance spectroscopy (IS), scanning electron microscopy (SEM), atomic force microscopy (AFM) and infra-red spectroscopy (IR). By using streptavidin labeled magnetic particles it was possible to functionalize the NPs assembly with biotin-Fab fragment K47 antibody. The designed biosensor had been successfully applied in rapid, simple, and accurate measurements of atrazine concentrations, with a significantly low detection limit of 5 ng/ml.