Yi Wan

Graphene oxide sheet-mediated silver enhancement for application to electrochemical biosensors.

Functionalized graphene oxide (GO) sheets coupled with a signal amplification method based on the nanomaterial-promoted reduction of silver ions for the sensitive and selective detection of bacteria. This paper aims to develop an electrochemical route combined with GO sheet-mediated Ag enhancement for biological/chemical analyte detection. A linear relationship between the stripping response and the logarithm of the bacterial concentration was obtained using an electrochemical technique for concentrations ranging from 1.8 × 10(2) to 1.8 × 10(8) cfu mL(-1), with a slope of 15.28 and a correlation coefficient of 0.995. Dot blot assay was used as a conventional immunoassay method for comparison with the electrochemical method, as well as to observe the quality of the anti-sulfate-reducing bacteria (SRB) antibody (Ab) used in the immunosensor. The GO sheet-mediated silver enhancement holds great potential for the rapid analysis of protein, DNA, and pathogens.

Impedimetric immunosensor doped with reduced graphene sheets fabricated by controllable electrodeposition for the non-labelled detection of bacteria

A facile, sensitive and reliable impedimetric immunosensor doped with reduced graphene sheets (RGSs) and combined with a controllable electrodeposition technique was developed for the selective detection of marine pathogenic sulphate-reducing bacteria (SRB). The morphology of RGSs and the electrochemical properties of RGSs-doped chitosan (CS) nanocomposite film were investigated by atomic force microscopy, Fourier transform infrared spectroscopy, and cyclic voltammetry (CV). Electrochemical impedance spectroscopy and CV were used to verify the stepwise assembly of the sensor system. Faradic impedance spectroscopy for charge transfer for the redox probe Fe(CN)(6)(3-/4-) was done to determine SRB concentrations. The diameter of the Nyquist diagram that is equal to the charge-transfer resistance (R(ct)) increased with increasing SRB concentration. A linear relationship between R(ct) and SRB concentration was obtained in the SRB concentration range of 1.8×10(1) to 1.8×10(7) cfu/ml. The impedimetric biosensor gave a distinct response to SRB, but had no obvious response to Vibrio angillarum. It showed a high selectivity for the detection of the pathogen. Based on a combination of the biocompatibility of CS and good electrical conductivity of RGSs, a nanocomposite film with novel architecture was used to immobilize biological and chemical targets and to develop a new type of biosensor.

Direct immobilisation of antibodies on a bioinspired architecture as a sensing platform

A sensitive and selective immunosensor for the nonlabeled detection of sulfate-reducing bacteria (SRB) is constructed using a self-polymerised polydopamine film as the immobilisation platform. Self-polymerisation of dopamine is used as a powerful approach for applying multifunctional coatings onto the surface of a gold electrode. The polydopamine film is used not only as the immobilisation platform, but also as a cross-linker reagent for the immobilisation of the anti-SRB antibody. The polydopamine film is loaded with a high density of anti-SRB antibodies linked to the substrate to obtain high response signals. The formation and fabrication of the biosensor and the quantification of antibody anchoring are monitored, and SRB detection is performed by either quartz crystal microbalance (QCM) or electrochemical impedance spectroscopy (EIS). After modeling the impedance Nyquist plots of the SRB/anti-SRB/polydopamine/gold electrode for increasing concentrations of SRB, the electron transfer resistance (R(ct)) is used as a measure of immunocomplex binding. The R(ct) is correlated with the concentration of bacterial cells in the range of 1.8×10(2) to 1.8×10(6) CFU mL(-1); the detection limit is 50 CFU mL(-1). This work demonstrates a new immobilisation platform for the development of a sensitive and label-less impedimetric and piezoelectric immunosensor. This immunosensor may be broadly applied in clinical diagnoses and the monitoring of water environmental pollution. The method proposed is distinct in its ease of application, use of a simple protocol, and mild reaction conditions. These allow it to be applied to a wide variety of materials.

Impedimetric biosensor based on cell-mediated bioimprinted films for bacterial detection.

This work presents the synthesis of bacteria-mediated bioimprinted films for selective bacterial detection. Marine pathogen sulfate-reducing bacteria (SRB) were chosen as the template bacteria. Chitosan (CS) doped with reduced graphene sheets (RGSs) was electrodeposited on an indium tin oxide electrode, and the resulting RGSs-CS hybrid film served as a platform for bacterial attachment. The electrodeposition conditions were optimized to obtain RGSs-CS hybrid films with excellent electrochemical performance. A layer of nonconductive CS film was deposited to embed the pathogen, and acetone was used to wash away the bacterial templates. Electrochemical impedance spectroscopy was performed to characterize the stepwise modification process and monitor the SRB population. Faradic impedance measurements revealed that the charge transfer resistance (R(ct)) increased with increased SRB concentration. A linear relationship between ΔR(ct) and the logarithm of SRB concentration was obtained within the concentration range of 1.0×10(4)cfum L(-1) to 1.0×10(8)cfum L(-1). The impedimetric sensor showed good selectivity towards SRB based on size and shape. Hence, selectivity for bacterial detection can be improved if the bioimprinting technique is combined with other bio-recognition elements.

Vancomycin-functionalised Ag@TiO2 phototoxicity for bacteria.

This study reports on the synthesis of vancomycin (Van)-functionalised Ag@TiO(2) nanoparticles and their enhanced bactericidal activities. Van-Ag@TiO(2) nanoparticles were prepared by nanoparticle deposition and chemical cross-linking reactions. The catalysts showed high efficiency for the degradation of methylene blue under ultraviolet (UV) illumination. The photocatalytic inactivation of the sulphate-reducing bacteria, Desulfotomaculum, was also studied under UV light irradiation and in the dark using aqueous mixtures of Ag, Ag@SiO(2), Ag@TiO(2), and Van-Ag@TiO(2). The Van-Ag@TiO(2) nanoparticles showed a capacity to target Van-sensitive bacteria. They also effectively prevented bacterial cell growth through the functionalised nanoparticles under UV irradiation for 1h. To investigate the specificity of the catalyst phototoxicity, a Van-resistant bacteria, Vibrio anguillarum, was used as the negative control. The results indicated that Van-Ag@TiO(2) nanoparticles had a higher selective phototoxicity for Van-sensitive bacteria. Therefore, the antibiotic molecule-functionalised core-shell nanoparticles allow for selective photokilling of pathogenic bacteria.

Monitoring microbial populations of sulfate-reducing bacteria using an impedimetric immunosensor based on agglutination assay

An impedimetric immunosensor was fabricated for rapid and non-labeled detection of sulfate-reducing bacteria, Desulforibrio caledoiensis (SRB) by immobilizing lectin-Concanavalin A using an agglutination assay. The immobilization of lectin was conducted using amine coupling on the surface of a gold (Au) electrode assembled with 11-Mercaptoundecanoic acid. Electrochemical impedance spectroscopy (EIS) was used to verify the stepwise assembly of the sensor system. The work conditions of the impedimetric immunosensor, such as pH of the buffer solutions and the incubation time of lectin, were optimized. Faradic impedance spectra for charge transfer for the redox probe Fe(CN)(6)(3-/4-)were measured to determine SRB concentrations. The diameter of the Nyquist diagram that is equal to the charge-transfer resistance (R(ct)) increased with increasing SRB concentration. A linear relationship between R(ct) and SRB concentration was obtained in SRB concentration range of 1.8 to 1.8 x 10(7)cfu/ml. The variation of the SRB population during the growth process was also monitored using the impedimetric immunosensor. This approach has great potential for simple, low-cost, and time-saving monitoring of microbial populations.

Controlled drug release characteristics and enhanced antibacterial effect of graphene oxide–drug intercalated layered double hydroxide hybrid films

Novel graphene oxide–benzylpenicillin (BP) anion intercalated Mg–Al layered double hydroxide (GO–BP-LDH) hybrid films were prepared via a simple solvent evaporation process. The release behaviour of BP anions and the antibacterial activity have also been investigated. The release time can be tailored by adjusting the film composition and increased by decreasing the GO/BP-LDH mass ratio. By applying release kinetic models to the drug release processes, the mechanism of release of BP anions from the GO–BP-LDH hybrid film was found to follow the first-order model. In addition, the hybrid film exhibited an enhanced synergistic antibacterial effect compared with the single GO film, which can be attributed to the combination of the antibacterial activity of GO and BP anions released from the hybrid film system.

Determination of sulphate-reducing bacteria based on vancomycin-functionalised magnetic nanoparticles using a modification-free quartz crystal microbalance

A fast, sensitive and reliable quartz crystal microbalance (QCM) biosensor is described for the selective detection of the marine pathogenic sulphate-reducing bacterium (SRB), D. desulfotomaculum. Based on the amplification of the response of vancomycin-functionalized magnetic nanoparticles (Van-mNPs), under an external magnetic field, the bacteria-mNPs conjugates attach to the surface of an Au electrode. The QCM biosensor gave a distinct response to the vancomycin-sensitive, D. desulfotomaculum, but had no obvious response to the vancomycin-resistant bacterium, Vibrio anguillarum. The effects of the optimization conditions such as the incubation time and pH on the detection were also investigated, respectively. Optimised assays showed that the biosensor could obtain the best response with a 30 min incubation of the bacteria with the Van-mNPs. A linear relationship between the QCM response and the logarithm of the bacterial concentration was observed in the range of 1.8 x 10(4) to 1.8 x 10(7)cfu/ml. The sensor system has a potential for further applications and provides a facile and sample method for detection of pathogenic bacteria.

Lysozyme as a recognition element for monitoring of bacterial population.

Bacterial infections remain a significant challenge in biomedicine and environment safety. Increasing worldwide demand for point-of-care techniques and increasing concern on their safe development and use, require a simple and sensitive bioanalysis for pathogen detection. However, this goal is not yet achieved. A design for fluorescein isothiocyanate-labeled lysozyme (FITC-LYZ), which provides quantitative binding information for gram-positive bacteria, Micrococcus luteus, and detects pathogen concentration, is presented. The functional lysozyme is used not only as the pathogenic detection platform, but also as a tracking reagent for microbial population in antibacterial tests. A nonlinear relationship between the system response and the logarithm of the bacterial concentration was observed in the range of 1.2×10(2)-1.2×10(5) cfu mL(-1). The system has a potential for further applications and provides a facile and simple method for detection of pathogenic bacteria. Meanwhile, the fluorescein isothiocyanate -labeled lysozyme is also employed as the tracking agent for antibacterial dynamic assay, which show a similar dynamic curve compared with UV-vis test.

A selective near-infrared fluorescent probe for hydrogen sulfide and its application in sulfate-reducing bacteria detection

Since sulfide has been considered as an important gasotransmitter in human activities, increased attention has been paid to its rapid detection. This work presented a novel near-infrared (NIR) fluorescent probe NIR-HS for sulfide detection based on thiolysis of dinitrophenyl ether. The probe was selective to sulfide over other common anion interferents with a large turn-on NIR fluorescence signal at 715 nm. Furthermore, the proposed NIR fluorescent probe NIR-HS was successfully applied in imaging and detection of sulfate-reducing bacteria, which could metabolize sulfide during their growth process, indicating that the probe NIR-HS was cell membrane permeable and could be used for sulfide monitoring in live cells and real samples.