Search

Electron transfer flavoprotein subunit beta (ETFB) of Leptospira interrogans is a biomarker for diagnosing leptospiral infection. Thus, the ETFB-specific nuclease-resistant RNA aptamer ETFB3-63 was developed and used in an electrochemical aptasensor to assay ETFB. Although the majority of reported biosensors detect various genes and antibodies of L. interrogans, this is the first attempt to construct an electrochemical biosensor to detect ETFB protein for the diagnosis of leptospiral infection. The ETFB protein can be detected without any extraction phase. In this assay, a single-stranded DNA probe complementary to the ETFB3-63 sequence was immobilized on a screen-printed carbon electrode (SPCE). The aptamer was then incubated and hybridized with the antisense probe on the SPCE. In the presence of ETFB, the aptamer dissociates from the aptamer/probe complex on the SPCE to bind with the protein. Methylene blue was then added to intercalate with the remaining hybridized aptamers, and its signal was measured using differential pulse voltammetry. The signal arising from the intercalated methylene blue decreased with increasing concentration of ETFB, showing a linear response in the range of 50–500 nM of ETFB and 10 to 109 leptospira cells per mL, respectively. The aptasensor signal was also specific to L. interrogans but not to 12 related bacteria tested. In addition, the aptasensor showed similar performance in detecting ETFB spiked in human serum to that in buffer, indicating that proteins in the serum do not interfere with the assay. Therefore, this assay has great potential to develop into a point-of-care electrochemical device that is accurate, cost-effective, and user-friendly for leptospirosis diagnosis. [ABSTRACT FROM AUTHOR]

In view of the presence of pathogenic Vibrio cholerae (V. cholerae) bacteria in environmental waters, including drinking water, which may pose a potential health risk to humans, an ultrasensitive electrochemical DNA biosensor for rapid detection of V. cholerae DNA in the environmental sample was developed. Silica nanospheres were functionalized with 3-aminopropyltriethoxysilane (APTS) for effective immobilization of the capture probe, and gold nanoparticles were used for acceleration of electron transfer to the electrode surface. The aminated capture probe was immobilized onto the Si-Au nanocomposite-modified carbon screen printed electrode (Si-Au-SPE) via an imine covalent bond with glutaraldehyde (GA), which served as the bifunctional cross-linking agent. The targeted DNA sequence of V. cholerae was monitored via a sandwich DNA hybridization strategy with a pair of DNA probes, which included the capture probe and reporter probe that flanked the complementary DNA (cDNA), and evaluated by differential pulse voltammetry (DPV) in the presence of an anthraquninone redox label. Under optimum sandwich hybridization conditions, the voltammetric genosensor could detect the targeted V. cholerae gene from 1.0 × 10−17–1.0 × 10−7 M cDNA with a limit of detection (LOD) of 1.25 × 10−18 M (i.e., 1.1513 × 10−13 µg/µL) and long-term stability of the DNA biosensor up to 55 days. The electrochemical DNA biosensor was capable of giving a reproducible DPV signal with a relative standard deviation (RSD) of <5.0% (n = 5). Satisfactory recoveries of V. cholerae cDNA concentration from different bacterial strains, river water, and cabbage samples were obtained between 96.5% and 101.6% with the proposed DNA sandwich biosensing procedure. The V. cholerae DNA concentrations determined by the sandwich-type electrochemical genosensor in the environmental samples were correlated to the number of bacterial colonies obtained from standard microbiological procedures (bacterial colony count reference method). [ABSTRACT FROM AUTHOR]

An essential biomarker for the early detection of cardiovascular diseases is serum homocysteine (Hcy). In this study, a molecularly imprinted polymer (MIP) and nanocomposite were used to create a label-free electrochemical biosensor for reliable Hcy detection. A novel Hcy-specific MIP (Hcy-MIP) was synthesized using methacrylic acid (MAA) in the presence of trimethylolpropane trimethacrylate (TRIM). The Hcy-MIP biosensor was fabricated by overlaying the mixture of Hcy-MIP and the carbon nanotube/chitosan/ionic liquid compound (CNT/CS/IL) nanocomposite on the surface of a screen-printed carbon electrode (SPCE). It showed high sensitivity, with a linear response of 5.0 to 150 µM (R2 of 0.9753) and with a limit of detection (LOD) at 1.2 µM. It demonstrated low cross-reactivity with ascorbic acid, cysteine, and methionine. Recoveries of 91.10–95.83% were achieved when the Hcy-MIP biosensor was used for Hcy at 50–150 µM concentrations. The repeatability and reproducibility of the biosensor at the Hcy concentrations of 5.0 and 150 µM were very good, with coefficients of variation at 2.27–3.50% and 3.42–4.22%, respectively. This novel biosensor offers a new and effective method for Hcy assay compared with the chemiluminescent microparticle immunoassay at the correlation coefficient (R2) of 0.9946. [ABSTRACT FROM AUTHOR]

We report a simple and rapid method for the synthesis of fluorescent gallium oxyhydroxide (GaOOH) nanoparticles from liquid Ga by a probe sonication method in the presence of H2O2 as an oxidant. The aspect ratio of the GaOOH nanoparticles is determined by the concentration of H2O2 and solution pH, as well as the probe energy and sonication time. Further surface modification with cyclodextrin to achieve biocompatibility for potential biomedical applications is reported where an example of cell uptake and fluorescence imaging is shown. [ABSTRACT FROM AUTHOR]

Background: Wolbachia is an endosymbiont bacterium generally found in about 40% of insects, including mosquitoes, but it is absent in Aedes aegypti which is an important vector of several arboviral diseasesdengue, chikungunya, zika, and yellow fever. The evidence that Wolbachia trans-infected Ae. aegypti mosquitoes lost their vectorial competence and became less capable of transmitting arboviruses to human hosts highlights the potential of using Wolbachia-based approaches for prevention and control of arboviral diseases. Recently, release of Wolbachia trans-infected Ae. aegypti has been deployed widely in many countries for the control of mosquito-borne viral diseases. Field surveillance and monitoring of Wolbachia presence in released mosquitoes is important for the success of these control programs. So far, a number of studies have reported the development of loop mediated isothermal amplification (LAMP) assays to detect Wolbachia in mosquitoes, but the methods still have some specificity and cost issues. Methodology/Principal findings: We describe here the development of a LAMP assay combined with the DNA strand displacement-based electrochemical sensor (BIOSENSOR) method to detect wAlbB Wolbachia in trans-infected Ae. aegypti. Our developed LAMP primers used a low-cost dye detecting system and 4 oligo nucleotide primers which can reduce the cost of analysis while the specificity is comparable to the previous methods. The detection capacity of our LAMP technique was 1.4 nM and the detection limit reduced to 2.2 fM when combined with the BIOSENSOR. Our study demonstrates that a BIOSENSOR can also be applied as a stand-alone method for detecting Wolbachia; and it showed high sensitivity when used with the crude DNA extracts of macerated mosquito samples without DNA purification. Conclusions/Significance: Our results suggest that both LAMP and BIOSENSOR, either used in combination or stand-alone, are robust and sensitive. The methods have good potential for routine detection of Wolbachia in mosquitoes during field surveillance and monitoring of Wolbachia-based release programs, especially in countries with limited resources. Author summary: Mosquito-borne diseases such as dengue, chikungunya, zika, and yellow fever are transmitted to humans mainly by the bites of Aedes aegypti mosquitoes. Controlling the vectors of these diseases relies mostly on the use of insecticides. However, the efficiency has been reduced through the development of insecticide resistance in mosquitoes. Wolbachia is an endosymbiotic bacterium that is naturally found in 40% of insects, including mosquitoes. The bacterium can protect its insect hosts from viral infections and can also cause sterility in insect host populations, therefore, providing an opportunity to use it for human disease control. Application of a Wolbachia trans-infected mosquitoes needs simple, rapid and sensitive methods for detecting the bacteria in released mosquitoes. In this paper, we develop the methods of LAMP and BIOSENSORS for detecting wAlbB Wolbachia in mosquitoes. Our positive LAMP reaction can be visualized by color change from violet to blue at a sensitivity of ≥ 10 pg of genomic DNA. When used in combination with the BIOSENSOR method, the sensitivity increases a millionfold without losing specificity. Our study suggests that both developed methods, either used in combination or stand-alone, are efficient and cost-effective, hence, they could be applied for routine surveys of Wolbachia in mosquito control programs that use Wolbachia-based approaches. [ABSTRACT FROM AUTHOR]

A tryptophan (Trp) sensor was investigated based on electrochemical impedance spectroscopy (EIS) of a molecularly imprinted polymer on a lysozyme amyloid fibril (MIP-AF). The MIP-AF was composed of aniline as a monomer chemically polymerized in the presence of a Trp template molecule onto the AF surface. After extracting the template molecule, the MIP-AF had cavities with a high affinity for the Trp molecules. The obtained MIP-AF demonstrated rapid Trp adsorption and substantial binding capacity (50 µM mg−1). Trp determination was studied using non-Faradaic EIS by drop drying the MIP-AF on the working electrode of a screen-printed electrode. The MIP-AF provided a large linear range (10 pM–80 µM), a low detection limit (8 pM), and high selectivity for Trp determination. Furthermore, the proposed method also indicates that the MIP-AF can be used to determine Trp in real samples such as milk and cancer cell media. [ABSTRACT FROM AUTHOR]

Infection with high-risk human papillomavirus (HPV) is a major risk factor for oral and cervical cancers. Hence, we developed a multianalyte electrochemical DNA biosensor that could be used for both oral and cervical samples to detect the high-risk HPV genotypes 16 and 18. The assay involves the sandwich hybridization of the HPV target to the silica-redox dye reporter probe and capture probe, followed by electrochemical detection. The sensor was found to be highly specific and sensitive, with a detection limit of 22 fM for HPV-16 and 20 fM for HPV-18, between the range of 1 fM and 1 µM. Evaluation with oral and cervical samples showed that the biosensor result was consistent with the nested PCR/gel electrophoresis detection. The biosensor assay could be completed within 90 min. Due to its simplicity, rapidity, and high sensitivity, this biosensor could be used as an alternative method for HPV detection in clinical laboratories as well as for epidemiological studies. [ABSTRACT FROM AUTHOR]

The ability of the technique of large-amplitude Fourier transformed (FT) ac voltammetry to facilitate the quantitative evaluation of electrode processes involving electron transfer and catalytically coupled chemical reactions has been evaluated. Predictions derived on the basis of detailed simulations imply that the rate of electron transfer is crucial, as confirmed by studies on the ferrocenemethanol (FcMeOH)-mediated electrocatalytic oxidation of ascorbic acid. Thus, at glassy carbon, gold, and boron-doped diamond electrodes, the introduction of the coupled electrocatalytic reaction, while producing significantly enhanced dc currents, does not affect the ac harmonics. This outcome is as expected if the [FcMeOH.sup.o/+] process remains fully reversible in the presence of ascorbic acid. In contrast, the ac harmonic components available from FT-ac voltammetry are predicted to be highly sensitive to the homogeneous kinetics when an electrocatalytic reaction is coupled to a quasi-reversible electron-transfer process. The required quasi-reversible scenario is available at an indium tin oxide electrode. Consequently, reversible potential, heterogeneous charge-transfer rate constant, and charge-transfer coefficient values of 0.19 V vs Ag/AgCl, 0.006 cm [s.sup.-1] and 0.55, respectively, along with a second-order homogeneous chemical rate constant of 2500 [M.sup.-1] [s.sup.-1] for the rate-determining step in the catalytic reaction were determined by comparison of simulated responses and experimental voltammograms derived from the dc and first to fourth ac harmonic components generated at an indium fin oxide electrode. The theoretical concepts derived for large-amplitude FT ac voltammetry are believed to be applicable to a wide range of important solution-based mediated electrocatalytic reactions.

We report a highly sensitive and selective multiplex assay by empowering an electrochemical DNA sensor with isothermal rolling circle amplification. The assay could simultaneously detect and discriminate three common entero-pathogens in a single reaction, with femtomolar sensitivity. It is useful for field- or resource-limited settings. [ABSTRACT FROM AUTHOR]

The heterogeneous rate constant (k) for the reduction of Fe(CN)63- at different UV-illuminated TiO2 polymorphs was determined by (1) cyclic voltammetry (CV) where a TiO2-modified fluorine-doped tin oxide (FTO)-coated glass was the working electrode, and (2) SECM approach curves at the same electrodes at open circuit. The combination of Fe(CN)63- with photogenerated electrons was found to be the rate determining step. The magnitude of the SECM values of k were in the order: anatase-rutile > anatase > rutile > amorphous, which agrees with the generally accepted photocatalytic efficiencies of the TiO2 polymorphs. In contrast, the CV-determined values of k followed the sequence: amorphous > rutile > anatase-rutile > anatase and were approximately one order of magnitude lower than the SECM-determined values. We suggest this indicates the CV determination of rate constants at photoactive films is problematic, due to factors such as reaction at the underlying electrode and the uncompensated resistance of the film. [ABSTRACT FROM AUTHOR]

Gold nanostructures were fabricated on a transparent indium tin oxide (ITO) coated PET substrate by an electrodeposition technique from a potassium gold (III) chloride solution for two different types of applications. It was found that the optical transparency of lightweight ITO electrodes could be maintained by depositing isolated gold nanostructures while opening up the use of these electrodes for inner sphere electron reactions, such as hydroquinone oxidation, which are not possible at ITO electrodes. For practical applications the adhesion of gold to the ITO electrode was improved by modifying the ITO surface with 3‐mercaptopropyl‐trimethoxysilane (MPS). Compared to Au/ITO, the Au/MPS/ITO electrode showed vastly improved electrochemical activity toward various electron transfer reactions when subjected to mechanical stress. The biosensing properties of the Au/MPS/ITO electrode was also investigated by studying the detection of immobilized DNA on the Au/MPS/ITO electrode via electrochemical impedance spectroscopy (EIS). [ABSTRACT FROM AUTHOR]

Room temperature liquid metals based on gallium demonstrate interesting physical and chemical properties when placed in electrochemical environments. In this work we explore the applicability of hanging liquid metal droplets as electrode materials for simple surface insensitive and surface sensitive electron transfer reactions, namely the electrochemical reduction of [Ru(NH 3 ) 6 ] 3+ and [Fe(CN) 6 ] 3− ions in solution. Significantly we found that for both redox species the electron transfer process was impeded at galinstan (68.5% Ga, 22.5% In and 10% Sn). This is related to the chemical reactivity of the Ga component which is oxidised when exposed to aqueous solutions of [Ru(NH 3 ) 6 ] 3+ and [Fe(CN) 6 ] 3− to generate the reduced form of the redox mediator. For the former this results in the production of Ru-red and Ru-brown cationic trimers in solution as well as gallium oxide on the surface of the liquid metal, and in the latter case the formation of solid Prussian Blue. [ABSTRACT FROM AUTHOR]

Abstract: In this work, the underpotential deposition of metals such as copper and lead during the electrochemical deposition of gold is investigated to understand the influence that the incorporation of a second metal has on the morphology of gold nanostructures. The incorporation of Pb or Cu, even at concentrations as low as 0.2 %, significantly influences the morphology of the deposit, where the formation of elongated structures from a central gold structure is favored. These nanostructures are characterized by cyclic voltammetry, X‐ray diffraction, scanning electron microscopy, and energy‐dispersive X‐ray spectroscopy and are tested for their suitability as a sensing layer for the electrochemical detection of nitrite ions in aqueous solution. A limit of detection of 0.3 μM is achieved with a linear range up to 1 mM, which is adequate for the determination of nitrite contained within food products. [ABSTRACT FROM AUTHOR]

Infectious diseases in goats, CAE in particular have widely affected the productivity of this animal and greatly affected the farmers and the small ruminant industry. Molecular technique such as LAMP has been applied to detect the CAEV proviral DNA specifically and sensitively but this technique has its own drawback. Gold is the most widely used nanoparticle (NP) which has excellent properties such as high surface area and compatibility with biomolecules. In this study, gold nanoparticle (AuNP) conjugated with modified oligonucleotides or gold nanoprobe (AuPr) was used to detect CAEV proviral DNA in LAMP product. The hybridization of AuPr to a complementary sequence on the LAMP product made the gold resistant to high salt concentration. This resistance or non-resistance of the AuNP is observed as color change in the mixture. This study, reported a simple method of visual detection of CAEV proviral DNA in LAMP product using AuPr. [ABSTRACT FROM AUTHOR]

DNA strand displacement is an attractive, enzyme-free target hybridization strategy for nano-biosensing. The target DNA induces a strand displacement reaction by replacing the pre-hybridized strand that is labeled with gold nanoparticles (AuNPs). Thus, the amount of displaced-AuNP-labeled strand is proportional to the amount of target DNA in the sample. The use of a magnetogenosensing technique to isolate the target DNA allows for a simple, one-pot detection approach, which minimizes possible carry-over contamination and pipetting errors. We sought a proof-of-concept for this technology in its ability to detect DNA-equivalent of hepatitis E virus (HEV), which causes acute viral hepatitis for which rapid and simple diagnostic methods remain limited. Signal detection was done via visual observation, spectrophotometry, and electrochemistry. The sensor demonstrated good sensitivity with detection limits of 10 pM (visual), 10 pM (spectrophotometry) and 1 fM (electrochemical). This sensor also exhibited high specificity for real target amplicons and could discriminate between perfect and mismatched sequences. Lyophilized biosensor reagents stored at 4 °C, 25 °C, and outdoor ambient temperature, were stable for up to 90, 50, and 40 days, respectively. The integration of magnetic separation and target DNA-induced strand displacement reaction in a dry reagent form makes the sensing platform easy-to-use and suitable for field settings. An electrochemical DNA sensor for the hepatitis E virus (HEV) which infects animals, transmits into food and contaminates water. Image 1 • One-pot detection with femtomolar limit of detection. • Signal interpretation can be achieved in three ways, visual observation, spectrophotometry, and electrochemical detection. • The integration of lyophilization and magnetogenosensing makes the test stable and easy to transport. • This platform can potentially detect nucleic acid of any viral or bacterial pathogen in infected samples. [ABSTRACT FROM AUTHOR]

The ability of a diagnostic test to detect multiple pathogens simultaneously is useful to obtain meaningful information for clinical treatment and preventive measures. We report a highly sensitive and specific electrochemical biosensor assay for simultaneous detection of three gene targets using quantum dots (QDs). The targets are novel non-protein coding RNA (npcRNA) sequences of Vibrio cholerae , Salmonella sp. and Shigella sp., which cause diarrheal diseases. QDs (PbS, CdS, ZnS) were synthesized and functionalized with DNA probes that were specific to each pathogen. Electrochemical detection of QDs was performed using square wave anodic stripping voltammetry (SWASV). The QDs gave distinct peaks at 0.5 V (PbS), 0.75 V (CdS) and 1.1 V (ZnS). There was no interference in signal response when all three QDs were mixed and detected simultaneously. The detection limits of single and multiplex assays with linear targets and PCR products were in the attomolar ranges. The high assay sensitivity, in combination with specific npcRNA sequences as novel diagnostic targets, makes it a viable tool for detecting pathogens from food, environment and clinical samples. [ABSTRACT FROM AUTHOR]