Gas Biosensor Arrays Based on Single-Stranded DNA-Functionalized Single-Walled Carbon Nanotubes for the Detection of Volatile Organic Compound Biomarkers Released by Huanglongbing Disease-Infected Citrus Trees.

Volatile organic compounds (VOCs) released by plants are closely associated with plant metabolism and can serve as biomarkers for disease diagnosis. Huanglongbing (HLB), also known as citrus greening or yellow shoot disease, is a lethal threat to the multi-billion-dollar citrus industry. Early detection of HLB is vital for removal of susceptible citrus trees and containment of the disease. Gas sensors are applied to monitor the air quality or toxic gases owing to their low-cost fabrication, smooth operation, and possible miniaturization. Here, we report on the development, characterization, and application of electrical biosensor arrays based on single-walled carbon nanotubes (SWNTs) decorated with single-stranded DNA (ssDNA) for the detection of four VOCs-ethylhexanol, linalool, tetradecene, and phenylacetaldehyde-that serve as secondary biomarkers for detection of infected citrus trees during the asymptomatic stage. SWNTs were noncovalently functionalized with ssDNA using π-π interaction between the nucleotide and sidewall of SWNTs. The resulting ssDNA-SWNT hybrid structure and device properties were investigated using Raman spectroscopy, ultraviolet (UV) spectroscopy, and electrical measurements. To monitor changes in the four VOCs, gas biosensor arrays consisting of bare SWNTs before and after being decorated with different ssDNA were employed to determine the different concentrations of the four VOCs. The data was processed using principal component analysis (PCA) and neural net fitting (NNF).

An oligonucleotide-functionalized carbon nanotube chemiresistor for sensitive detection of mercury in saliva.

Divalent mercuric (Hg(2+)) ion and monomethyl mercury (CH3Hg(+)) are two forms of mercury that are known to be highly toxic to humans. In this work, we present a highly selective, sensitive and label-free chemiresistive biosensor for the detection of both, Hg(2+) and CH3Hg(+) ions using DNA-functionalized single-walled carbon nanotubes (SWNTs). The SWNTs were functionalized with the capture oligonucleotide, polyT, using a linker molecule. The polyT was hybridized with polyA to form a polyT:polyA duplex. Upon exposure to mercury ions, the polyT:polyA duplex dehybridizes and a T-Hg(2+)-T duplex is formed. This structure switch leads to the release of polyA from the SWNT surface and correspondingly a change in the resistance of the chemiresistive biosensor is observed, which is used to quantify the mercury ion concentration. The biosensor showed a wide dynamic range of 0.5 to 100 nM for the detection of CH3Hg(+) ions in buffer solution with a sensitivity of 28.34% per log (nM) of CH3Hg(+). Finally, real world application of the biosensor was demonstrated by the detection of Hg(2+) and CH3Hg(+) ions in simulated saliva samples spiked with a known concentration of mercury ions.

Electrochemically Functionalized Seamless Three-Dimensional Graphene-Carbon Nanotube Hybrid for Direct Electron Transfer of Glucose Oxidase and Bioelectrocatalysis.

Three-dimensional seamless chemical vapor deposition (CVD) grown graphene-carbon nanotubes (G-CNT) hybrid film has been studied for its potential in achieving direct electron transfer (DET) of glucose oxidase (GOx) and its bioelectrocatalytic activity in glucose detection. A two-step CVD method was employed for the synthesis of seamless G-CNT hybrid film where CNTs are grown on already grown graphene film on copper foil using iron as a catalyst. Physical characterization using SEM and TEM show uniform dense coverage of multiwall carbon nanotubes (MWCNT) grown directly on graphene with seamless contacts. The G-CNT hybrid film was electrochemically modified to introduce oxygenated functional groups for DET favorable immobilization of GOx. Pristine and electrochemically functionalized G-CNT film was characterized by electrochemical impedance spectroscopy (EIS), cyclic voltammetry, X-ray photoelectron-spectroscopy, and Raman spectroscopy. The DET between GOx and electrochemically oxidized G-CNT electrode was studied using cyclic voltammetry which showed a pair of well-defined and quasi-reversible redox peaks with a formal potential of -459 mV at pH 7 corresponding to the redox site of GOx. The constructed electrode detected glucose concentration over the clinically relevant range of 2-8 mM with the highest sensitivity of 19.31 µA/mM/cm(2) compared to reported composite hybrid electrodes of graphene oxide and CNTs. Electrochemically functionalized CVD grown seamless G-CNT structure used in this work has potential to be used for development of artificial mediatorless redox enzyme based biosensors and biofuel cells.

Label-Free Electrical Immunosensor for Highly Sensitive and Specific Detection of Microcystin-LR in Water Samples.

Microcystin-LR (MCLR) is one of the most commonly detected and toxic cyclic heptapeptide cyanotoxins released by cyanobacterial blooms in surface waters, for which sensitive and specific detection methods are necessary to carry out its recognition and quantification. Here, we present a single-walled carbon nanotube (SWCNTs)-based label-free chemiresistive immunosensor for highly sensitive and specific detection of MCLR in different source waters. MCLR was initially immobilized on SWCNTs modified interdigitated electrode, followed by incubation with monoclonal anti-MCLR antibody. The competitive binding of MCLR in sample solutions induced departure of the antibody from the antibody-antigen complexes formed on SWCNTs, resulting in change in the conductivity between source and drain of the sensor. The displacement assay greatly improved the sensitivity of the sensor compared with direct immunoassay on the same device. The immunosensor exhibited a wide linear response to log value of MCLR concentration ranging from 1 to 1000 ng/L, with a detection limit of 0.6 ng/L. This method showed good reproducibility, stability and recovery. The proposed method provides a powerful tool for rapid and sensitive monitoring of MCLR in environmental samples.

Electronic detection of microRNA at attomolar level with high specificity.

Small RNA (19-23 nucleotides) molecules play an important role in gene regulation, embryonic differentiation, hematopoiesis, and a variety of cancers. Here, we present an ultrasensitive, extremely specific, label-free, and rapid electronic detection of microRNAs (miRNAs) using a carbon nanotubes field-effect transistor functionalized with the Carnation Italian ringspot virus p19 protein biosensor. miRNA-122a was chosen as the target, which was first hybridized to a probe molecule. The probe-miRNA duplex was then quantified by measuring the change in resistance of biosensor resulting from its binding to p19, which selects 21-23 bp RNA duplexes in a size-dependent but sequence-independent manner. The biosensor displayed a wide dynamic range up to 10(-14) M and was able to detect as low as 1 aM miRNA in the presence of a million-fold excess of total RNA, paving the way for simple, point-of-care, low-cost early detection of miRNA as a biomarker in diagnosis of many diseases, including cancer.

Asymptomatic Diagnosis of Huanglongbing Disease Using Metalloporphyrin Functionalized Single-Walled Carbon Nanotubes Sensor Arrays.

Porphyrins, with or without metal ions (MPs), have been explored and applied in optical and electrochemical sensor fields owing to their special physicochemical properties. The presence of four nitrogen atoms at the centers of porphyrins means that porphyrins chelate most metal ions, which changes the binding ability of MPs with gas molecules via non-specific binding. In this article, we report hybrid chemiresistor sensor arrays based on single-walled carbon nanotubes (SWNTs) non-covalently functionalized with six different MPs using the solvent casting technique. The characteristics of MP-SWNTs were investigated through various optical and electrochemical methods, including UV spectroscopy, Raman, atomic force microscopy, current-voltage (I-V), and field-effect transistor (FET) measurement. The proposed sensor arrays were employed to monitor the four VOCs (tetradecene, linalool, phenylacetaldehyde, and ethylhexanol) emitted by citrus trees infected with Huanglongbing (HLB), of which the contents changed dramatically at the asymptomatic stage. The sensitivity to VOCs could change significantly, exceeding the lower limits of the SWNT-based sensors. For qualitative and quantitative analysis of the four VOCs, the data collected by the sensor arrays were processed using different regression models including partial least squares (PLS) and an artificial neural network (ANN), which further offered a diagnostic basis for Huanglongbing disease at the asymptomatic stage.

Graphene nanogap electrodes in electrical biosensing.

Graphene nanogap electrodes are reported here for the first time in an electrical biosensor for the detection of biomolecular interactions. Streptavidin-biotin was chosen as a model system for evaluating the sensor's performance. High-affinity interactions of streptavidin-gold nanoparticles (strep-AuNPs) to the biotin-functionalized nanogap localizes AuNPs, thereby bridging the gap and resulting in changes in device conductance. Biosensing performance was optimized by varying the gap size, AuNP diameter, and streptavidin coverage on AuNPs. The sensitivity and limit of detection (LOD) of streptavidin detection with the optimized parameters were determined to be 0.3 µA/nM and 0.25 pM, respectively. The proposed platform suggests high potential as a portable point-of-use biosensor for the detection of other affinity-based biomolecular interactions, such as antigen-antibody, nucleic acid, or chemo-selective interactions.

Monitoring of microbial cell viability using nanostructured electrodes modified with Graphene/Alumina nanocomposite.

Microbial infections are rapidly increasing; however most of the existing microbiological and molecular detection methods are time consuming and/or cannot differentiate between the viable and dead cells which may overestimate the risk of infections. Therefore, a bioelectrochemical sensing platform with a high potential to the microbial-electrode interactions was designed based on decorated graphene oxide (GO) sheet with alumina (Al2O3) nanocrystals. GO-Al2O3 nanocomposite was synthesized using self-assembly of GO and Al2O3 and characterized using the scanning electron microscopy (SEM), transmission electron microscopy (TEM), x-ray diffraction (XRD), Raman-spectroscopy, electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV). Enhancement of electrocatalytic activity of the composite-modified electrode was demonstrated. Thus, using the GO-Al2O3 nanocomposite modified electrode, the cell viability was determined by monitoring the bioelectrochemical response of the living microbial cells (bacteria and yeast) upon stimulation with carbon source. The bioelectrochemical assay was optimized to obtain high sensitivity and the method was applied to monitor cell viability and screen susceptibility of metabolically active cells (E. coli, B. subtilis, Enterococcus, P. aeruginosa and Salmonella typhi) to antibiotics such as ampicillin and kanamycin. Therefore, the developed assay is suitable for cell proliferation and cytotoxicity testing.

Carbon nanomaterial-based electrochemical biosensors for label-free sensing of environmental pollutants.

Carbon allotropes such as graphene and carbon nanotubes, have been incorporated in electrochemical biosensors for highly sensitive and selective detection of various analytes. The superior physical and electrical properties like high carrier mobility, ambipolar electric field effect, high surface area, flexibility and their compatibility with microfabrication techniques makes these carbon nanomaterials easy to integrate in field-effect transistor (FET)/chemiresistor type configuration which is suitable for portable and point-of-use/field-deployable sensors. This review covers the synthesis of carbon nanostructures (graphene and CNTs) and their integration into devices using various fabrication methods. Finally, we discuss the recent reports showing different sensing platforms that incorporate biomolecules like enzymes, antibodies and aptamers as recognition elements for fabrication of simple, low cost, compact biosensors that can be used for on-site, rapid environmental monitoring of environmental pollutants like pathogens, heavy metals, pesticides and explosives.

Thermal conductivity of graphene with defects induced by electron beam irradiation.

We investigate the thermal conductivity of suspended graphene as a function of the density of defects, ND, introduced in a controllable way. High-quality graphene layers are synthesized using chemical vapor deposition, transferred onto a transmission electron microscopy grid, and suspended over ∼7.5 µm size square holes. Defects are induced by irradiation of graphene with the low-energy electron beam (20 keV) and quantified by the Raman D-to-G peak intensity ratio. As the defect density changes from 2.0 × 10(10) cm(-2) to 1.8 × 10(11) cm(-2) the thermal conductivity decreases from ∼(1.8 ± 0.2) × 10(3) W mK(-1) to ∼(4.0 ± 0.2) × 10(2) W mK(-1) near room temperature. At higher defect densities, the thermal conductivity reveals an intriguing saturation-type behavior at a relatively high value of ∼400 W mK(-1). The thermal conductivity dependence on the defect density is analyzed using the Boltzmann transport equation and molecular dynamics simulations. The results are important for understanding phonon - point defect scattering in two-dimensional systems and for practical applications of graphene in thermal management.