Nanostructured Photoelectrochemical Biosensing Platform for Cancer Biomarker Detection

The innovative photoelectrochemical properties of multifunctional nanomaterials are here investigated for the development of biosensing platforms for rapid and sensitive detection of a class of cancer biomarker candidates, known as microRNAs. Many different transducers have been proposed, so far, for microRNA detection. Recently, with the emergence of novel photoelectrochemically active species and new detection schemes, photoelectrochemistry has received increasing attention. Gold nanostructures have been, here, used to modify TiO2 electrodes. The surface of the nanostructured platform has been modified by nucleic acid capture probes (CPs). Biotinylated target miRNAs have been recognized by the specific CPs. The biosensing platform has been incubated with streptavidin alkaline phosphatase and exposed to a proper substrate. The product of the enzymatic reaction has been photoelectrochemically monitored. A compact and hand-held analytical device has been developed in order to have a final prototype in line with the concept of point of care testing. (C) 2017 The Authors. Published by Elsevier Ltd

Electrochemical Liposome-Based Biosensors for Nucleic Acid Detection

MicroRNAs (miRNAs) are intensely studied as candidates for diagnostic and prognostic clinical biomarkers. Faradic impedance spectroscopy (EIS) and differential pulse voltammetry (DPV), coupled to disposable gold electrodes and enzyme amplification of the hybridization event, were used for the development of biosensors for the detection of miRNAs. Biotin-labeled liposomes were employed as nanointerfaces that amplify the primary miRNA-sensing events by their association to the probe\u2013/DNA\u2013miRNA\u2013analyte complex generated onto the transducer

Strategies for the development of an electrochemical bioassay for TNF-alpha detection by using a non-immunoglobulin bioreceptor

TNF-alpha is an inflammatory cytokine produced by the immune system. Serum TNF-alpha level is elevated in some pathological states such as septic shock, graft rejection, HIV infection, neurodegenerative diseases, rheumatoid arthritis and cancer. Detecting trace amount of TNF-alpha is, also, very important for the understanding of tumor biological processes. Detection of this key biomarker is commonly achieved by use of ELISA or cytofluorimetric based methods. In this study the traditional optical detection was replaced by differential pulse voltammetry (DPV) and an affinity molecule, produced by evolutionary approaches, has been tested as capture bioreceptor. This molecule, namely a combinatorial non-immunoglobulin protein (Affibody (R)) interacts with TNF-alpha selectively and was here tested in a sandwich assay format. Moreover magnetic beads were used as support for bioreceptor immobilization and screen printed carbon electrodes were used as transducers.TNF-alpha calibration curve was performed, obtaining the detection limit of 38 pg/mL, the quantification range of 76-5000 pg/mL and RSD%=7. Preliminary results of serum samples analysis were also reported. (C) 2016 Elsevier B.V. All rights reserved

Glyphosate Determination by Coupling an Immuno-Magnetic Assay with Electrochemical Sensors

Glyphosate (N-(phosphonomethyl)glycine) is the most frequently used broad-spectrum herbicide worldwide. Its mechanism of action is based on the inhibition of an enzyme that is essential to plant growth. Its intensive use has caused global contamination to occur, which has not only affected the ecosystems, but even food and other objects of common use. Thus, there is a pronounced need for developing analytical methods for glyphosate determination in different matrices. Here, an electrochemical competitive immunoassay, based on the use of antibody-modified magnetic particles, has been developed. Tetramethylbenzidine (TMB) has been used as an enzymatic substrate. The extent of the affinity reaction has been achieved by monitoring the current value, due to the reduction of the enzymatic product. A disposable screen-printed electrochemical cell has been used. The calibration curve has been recorded in the 0–10,000 ng/L concentration range, with a detection limit of 5 ng/L and quantification limit of 30 ng/L. The electrochemical immunoassay has also been applied to the analysis of spiked beer samples

Trends and Perspectives in Immunosensors for Determination of Currently-Used Pesticides: The Case of Glyphosate, Organophosphates, and Neonicotinoids

Pesticides, due to their intensive use and their peculiar chemical features, can persist in the environment and enter the trophic chain, thus representing an environmental risk for the ecosystems and human health. Although there are several robust and reliable standard analytical techniques for their monitoring, the high frequency of contamination caused by pesticides requires methods for massive monitoring campaigns that are capable of rapidly detecting these compounds in many samples of different origin. Immunosensors represent a potential tool for simple, rapid, and sensitive monitoring of pesticides. Antibodies coupled to electrochemical or optical transducers have resulted in effective detection devices. In this review, the new trends in immunosensor development and the application of immunosensors for the detection of pesticides of environmental concern—such as glyphosate, organophosphates, and neonicotinoids—are described

Improving impedimetric nucleic acid detection by using enzyme-decorated liposomes and nanostructured screen-printed electrodes

Sensitive impedimetric detection of miR-222, a miRNA sequence found in many lung tumors, was investigated by using gold-nanostructured disposable carbon electrodes and enzyme-decorated liposomes. The proposed method was based on the immobilization of thiolated DNA capture probes onto gold-nanostructured carbon surfaces. Afterwards, the capture probes were allowed to hybridize to the target miRNAs. Finally, enzyme-decorated liposomes were used as labels to amplify the miRNA sensing, by their association with the probe-miRNA hybrids generated on the nanostructured transducer. By using this amplification route a limit of detection of 0.400 pM, a limit of quantification of 1.70 pM, and an assay range spanning three orders of magnitude (1.70-900 pM) were obtained (RSD % = 13). This limit of quantification was 20 times lower than that obtained using a simple enzyme conjugate for the detection. A comparison was also made with gold screen-printed transducers. In this case, a limit of quantification approximately 70 times lower was found by using the nanostructured transducers. Application of the optimized assay in serum samples was also demonstrated

Health and carcinogenic risk evaluation for cohorts exposed to PAHs in petrochemical workplaces in Rawalpindi city (Pakistan)

<p>This study presents the analyses of urinary biomarkers (1-OHPyr, α- and β-naphthols) of polycyclic aromatic hydrocarbons (PAHs) exposure and biomarkers of effect (i.e. blood parameters) in petroleum-refinery workers (RFs) and auto-repair workers (MCs). Exposed subjects had higher concentrations of white blood cell (WBC) count than control subjects (CN) subjects (5.31 × 10³ μL⁻¹ in exposed vs. 5.15 × 10³ μL⁻¹ in CN subjects), while the biomarker of oxidative DNA damage (8-OHdG) was significantly higher in MCs. The exposure among these two cohorts could be influenced by the ambience of the workplaces; in fact, MCs’ shops are relatively damp and enclosed workplaces in comparison with the indoor environment of refineries. PAHs in the dust samples from mechanical workshops probably originated from mixed sources (traffic exhaust and petroleum spills), while the incremental lifetime cancer risk (ILCR) for MCs showed moderate-to-low cancer risk from exposure to dust-bound PAHs. The study shows that increasing PAH exposure can be traced in MC workstations and needs to be investigated for the safety of public health.</p