Ultrasensitive SARS-CoV-2 diagnosis by CRISPR-based screen-printed carbon electrode
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ABSTRACT:
SUBMITTER: Wu L
PROVIDER: S-EPMC9249825 | biostudies-literature |
REPOSITORIES: biostudies-literature
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Project description:Herein, a novel molecularly imprinted polymer (MIP) based electrochemical sensor for the determination of the receptor-binding domain of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2-RBD) has been developed. For this purpose, first, a macroporous gold screen-printed electrode (MP-Au-SPE) has been fabricated. The MIP was then synthesized on the surface of the MP-Au-SPE through the electro-polymerization of ortho-phenylenediamine in the presence of SARS-CoV-2-RBD molecules as matrix polymer, and template molecules, respectively. During the fabrication process, the SARS-CoV-2-RBD molecules were embedded in the polymer matrix. Subsequently, the template molecules were removed from the electrode by using alkaline ethanol. The template molecules removal was studied using cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), scanning electron microscope (SEM), energy-dispersive X-ray spectroscopy (EDX), and attenuated total reflectance spectroscopy (ATR). The fabricated MIP film acted as an artificial recognition element for the measurement of SARS-CoV-2-RBD. The EIS technique was used for the measurement of the SARS-CoV-2-RBD in the saliva solution. The electron transfer resistance (Ret) of the MIP-based sensor in a ferri/ferrocyanide solution increased as the SARS-CoV-2-RBD concentration increased due to the occupation of the imprinted cavities by the SARS-CoV-2-RBD. The MIP-based sensor exhibited a good response to the SARS-CoV-2-RBD in the concentration range between 2.0 and 40.0 pg mL-1 with a limit of detection of 0.7 pg mL-1. The obtained results showed that the fabricated MIP sensor has high selectivity sensitivity, and stability.
| S-EPMC8547854 | biostudies-literature
Project description:Worldwide, human health is affected by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Hence, the fabrication of the biosensors to diagnose SARS-CoV-2 is critical. In this paper, we report an electrochemical impedance spectroscopy (EIS)-based aptasensor for the determination of the SARS-CoV-2 receptor-binding domain (SARS-CoV-2-RBD). For this purpose, the carbon nanofibers (CNFs) were first decorated with gold nanoparticles (AuNPs). Then, the surface of the carbon-based screen-printed electrode (CSPE) was modified with the CNF-AuNP nanocomposite (CSPE/CNF-AuNP). After that, the thiol-terminal aptamer probe was immobilized on the surface of the CSPE/CNF-AuNP. The surface coverage of the aptamer was calculated to be 52.8 pmol·cm-2. The CSPE/CNF-AuNP/Aptamer was then used for the measurement of SARS-CoV-2-RBD by using the EIS method. The obtained results indicate that the signal had a linear-logarithmic relationship in the range of 0.01-64 nM with a limit of detection of 7.0 pM. The proposed aptasensor had a good selectivity to SARS-CoV-2-RBD in the presence of human serum albumin; human immunoglobulins G, A, and M, hemagglutinin, and neuraminidase. The analytical performance of the aptasensor was studied in human saliva samples. The present study indicates a practical application of the CSPE/CNF-AuNP/Aptamer for the determination of SARS-CoV-2-RBD in human saliva samples with high sensitivity and accuracy.
| S-EPMC8945915 | biostudies-literature
Project description:Regulatory bodies play a crucial role in establishing limits for food additives to ensure food quality and safety of food products, as excessive usage poses risks to consumers. In the context of processed animal-based foodstuffs, nitrite is commonly utilized as a means to slow down bacterial degradation. In this study, we have successfully leveraged the redox activity of an electrochemically deposited polydopamine (pDA) film onto gold nanoparticle (AuNP)-modified screen-printed electrodes (SPCE) to develop a sensitive and versatile methodology for the detection of nitrite using redox capacitance spectroscopy. By exploiting the interaction of the AuNPs/pDA electroactive interface with the target nitrite ions, we observed distinct changes in the redox distribution, subsequently leading to modifications in the associated redox capacitance. This alteration enables the successful detection of nitrite, exhibiting a linear response within the concentration range of 10 to 500 μM, with a limit of detection of 1.98 μM (S/N = 3). Furthermore, we applied the developed sensor to analyze nitrite levels in processed meats, yielding good recoveries. These results demonstrate the potential of our approach as a promising method for routine detection of ions.
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Project description:To help meet the global demand for reliable and inexpensive COVID-19 testing and environmental analysis of SARS-CoV-2, the present work reports the development and application of a highly efficient disposable electrochemical immunosensor for the detection of SARS-CoV-2 in clinical and environmental matrices. The sensor developed is composed of a screen-printed electrode (SPE) array which was constructed using conductive carbon ink printed on polyethylene terephthalate (PET) substrate made from disposable soft drink bottles. The recognition site (Spike S1 Antibody (anti-SP Ab)) was covalently immobilized on the working electrode surface, which was effectively modified with carbon black (CB) and gold nanoparticles (AuNPs). The immunosensing material was subjected to a multi-technique characterization analysis using X-ray diffraction (XRD), transmission electron microscopy (TEM), and scanning electron microscopy (SEM) with elemental analysis via energy dispersive spectroscopy (EDS). The electrochemical characterization of the electrode surface and analytical measurements were performed using cyclic voltammetry (CV) and square-wave voltammetry (SWV). The immunosensor was easily applied for the conduct of rapid diagnoses or accurate quantitative environmental analyses by setting the incubation period to 10 min or 120 min. Under optimized conditions, the biosensor presented limits of detection (LODs) of 101 fg mL-1 and 46.2 fg mL-1 for 10 min and 120 min incubation periods, respectively; in addition, the sensor was successfully applied for SARS-CoV-2 detection and quantification in clinical and environmental samples. Considering the costs of all the raw materials required for manufacturing 200 units of the AuNP-CB/PET-SPE immunosensor, the production cost per unit is 0.29 USD.
| S-EPMC9998283 | biostudies-literature
Project description:Early diagnosis is highly crucial for life-saving and transmission management of tuberculosis (TB). Despite the low sensitivity and time-consuming issues, TB antigen detection still relies on conventional smear microscopy and culture techniques. To address this limitation, we report the development of the first amperometric dual aptasensor for the simultaneous detection of Mycobacterium tuberculosis secreted antigens CFP10 and MPT64 for better diagnosis and control of TB. The developed sensor was based on the aptamers-antibodies sandwich assay and detected by chronoamperometry through the electrocatalytic reaction between peroxidase-conjugated antibodies, H2O2, and hydroquinone. The CFP10 and MPT64 aptamers were immobilized via carbodiimide covalent chemistry over the disposable dual screen-printed carbon electrodes modified with a 4-carboxyphenyl diazonium salt. Under optimized conditions, the aptasensor achieved a detection limit of 1.68 ng mL-1 and 1.82 ng mL-1 for CFP10 and MPT64 antigens, respectively. The developed assay requires a small sample amount (5 µL) and can be easily performed within 2.5 h. Finally, the dual aptasensor was successfully applied to clinical sputum samples with the obtained diagnostic sensitivity (n = 24) and specificity (n = 13) of 100%, respectively, suggesting the readiness of the developed assay to be used for TB clinical application.
| S-EPMC9688523 | biostudies-literature
Project description:We report a facile strategy for developing reagentless amperometric pyruvate biosensors based on enzyme nanoparticles (EnNPs). The EnNPs were prepared using pyruvate oxidase crosslinked with graphene quantum dots. Before EnNP immobilization, screen-printed carbon electrodes (SPCEs) were modified with Prussian blue, a biocompatible coordination polymer. The biosensor system was optimized in terms of the working potential and pH value. At pH 7.0 in 50 mM phosphate-buffered solution, the biosensor showed optimal characteristics under an applied potential of -0.10 V versus an internal pseudo-Ag reference electrode. Using these optimized conditions, the biosensor performance was characterized via the chronoamperometric technique. The EnNP-immobilized SPCE exhibited a dynamic linear range from 10 to 100 μM for pyruvate solution, and a sensitivity of 40.8 μA mM-1 cm-2 was recorded. The observed detection limit of the biosensor was 0.91 μM (S/N = 3) and it showed strong anti-inference capability under the optimized working potential. Furthermore, the practical applicability of the proposed biosensor was studied in fish serum samples.
| S-EPMC7689939 | biostudies-literature
Project description:This work demonstrates the analysis of epinephrine (EP) and uric acid (UA) in a single drop (the volume of the test solution was only 50 µL) using a screen-printed carbon electrode (SPCE) sensor and square-wave voltammetry (SWV). The limit of detection, limit of quantification, linearity, accuracy, precision, and robustness were validated. The normality of the experimental data was tested and confirmed for both methods. Heteroscedasticity was checked by residual analysis followed by a statistical F-test. The latter was confirmed for both analytes. The low relative standard deviations (RSD) at all calibration points and repetitive slopes justified the use of a calibration curve; therefore, the standard addition methodology was avoided (the latter is common in electroanalysis, but time-consuming). Since the conditions for using an ordinary least squares (OLS) regression were not met, weighted linear regression (WLR) was used to improve the accuracy of the analytical results at low concentrations of the analytes. In this manner, the best weighted model was determined and used for the quantification. A comparison was made between the OLS and WLR methods to show the necessity of using the WLR method for EP and UA analysis. The newly developed and validated methods were also shown to be effective in the analysis of real samples. The content of EP in an EP auto-injector and UA in human urine was tested by employing the best weighted model. For EP and UA, the accuracy in terms of the average recovery value was 101.01% and 94.35%, and precision in terms of RSD was 5.65% and 2.75%, respectively. A new analytical methodology is presented that uses a low volume (a single drop), and it offers the advantage of electroanalysis for on-site analysis, where conventional chromatographic techniques cannot be easily employed. Furthermore, the developed technique has additional advantages in terms of speed, cost, and miniaturization.
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Project description:A original electrochemical sensing platform, based on screen-printed electrodes modification with plasma polymerized acrylonitrile (pp-AN) nanofilms is proposed. For that purpose, plasma-enhanced chemical vapor deposition (PECVD) process was conducted in a parallel plate (13.56 MHz) plasma reactor for 2 min with discharge power of 10 W. The surface topography and electrochemical properties of prepared sensors were investigated by X-ray photoelectron spectroscopy, scanning electron microscopy, energy dispersion spectroscopy, electrochemical impedance spectroscopy, and cyclic voltammetry. The electrochemical characteristics of pp-AN/SPCE and pp-AN/SPAuE sensors was investigated for model redox pair [Fe(CN)6]4-/3-. Conducted research confirmed the excellent chemical stability, durability, wide potential window, high signal-to-noise (S/N) ratio, and, most importantly, the ability to standardize the sensors. The pp-AN/SPCE sensor was applied to the determination of bupropion, an antidepressant drug whose intake has increased dramatically during the COVID-19 pandemic. The voltammetric response of pp-AN/SPCE for BUP was linear in two concentration ranges of 0.63-10.0 and 10.0-50.0 μmol L-1, with a detection limit of 0.21 μmol L-1. Satisfactory recoveries (96.2-102%) and good precision (RSD below 4.1%) obtained for environmental and biological samples confirmed the usefulness of the sensor for the analysis of various kinds of samples.
| S-EPMC10499721 | biostudies-literature
Project description:Simple, rapid and accurate detection of ethanol concentration in blood is very crucial in the diagnosis and management of potential acute ethanol intoxication patients. A novel electrochemical detection method was developed for the quantification of ethanol in human plasma with disposable unmodified screen-printed carbon electrode (SPCE) without sample preparation procedure. Ethanol was detected indirectly by the reaction product of ethanol dehydrogenase (ADH) and cofactor nicotinamide adenine dinucleotide (NAD(+)). Method validation indicated good quantitation precisions with intra-day and inter-day relative standard deviations of ?9.4% and 8.0%, respectively. Ethanol concentration in plasma is linear ranging from 0.10 to 3.20?mg/mL, and the detection limit is 40.0??g/mL (S/N?>?3). The method shows satisfactory correlation with the reference method of headspace gas chromatography in twenty human plasma samples (correlation coefficient 0.9311). The proposed method could be applied to diagnose acute ethanol toxicity or ethanol-related death.
| S-EPMC4804280 | biostudies-literature
Project description:Electrochemical sensors consisting of screen-printed electrodes (SPEs) are recurrent devices in the recent literature for applications in different fields of interest and contribute to the expanding electroanalytical chemistry field. This is due to inherent characteristics that can be better (or only) achieved with the use of SPEs, including miniaturization, cost reduction, lower sample consumption, compatibility with portable equipment, and disposability. SPEs are also quite versatile; they can be manufactured using different formulations of conductive inks and substrates, and are of varied designs. Naturally, the analytical performance of SPEs is directly affected by the quality of the material used for printing and modifying the electrodes. In this sense, the most varied carbon nanomaterials have been explored for the preparation and modification of SPEs, providing devices with an enhanced electrochemical response and greater sensitivity, in addition to functionalized surfaces that can immobilize biological agents for the manufacture of biosensors. Considering the relevance and timeliness of the topic, this review aimed to provide an overview of the current scenario of the use of carbonaceous nanomaterials in the context of making electrochemical SPE sensors, from which different approaches will be presented, exploring materials traditionally investigated in electrochemistry, such as graphene, carbon nanotubes, carbon black, and those more recently investigated for this (carbon quantum dots, graphitic carbon nitride, and biochar). Perspectives on the use and expansion of these devices are also considered.
| S-EPMC10136782 | biostudies-literature