Browse
Submit Data
Databases
API
Help

Dataset Information

16 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Ultrasensitive SARS-CoV-2 diagnosis by CRISPR-based screen-printed carbon electrode

ABSTRACT: Early and accurate diagnosis of SARS-CoV-2 was crucial for COVID-19 control and urgently required ultra-sensitive and rapid detection methods. CRISPR-based detection systems have great potential for rapid SARS-CoV-2 detection, but detecting ultra-low viral loads remains technically challenging. Here, we report an ultrasensitive CRISPR/Cas12a-based electrochemical detection system with an electrochemical biosensor, dubbed CRISPR-SPCE, in which the CRISPR ssDNA reporter was immobilized onto a screen-printed carbon electrode. Electrochemical signals are detected due to CRISPR cleavage, giving enhanced detection sensitivity. CRISPR-SPCE enables ultrasensitive SARS-CoV-2 detection, reaching as few as 0.27 copies μL−1. Moreover, CRISPR-SPCE is also highly specific and inexpensive, providing a fast and simple SARS-CoV-2 assay. Graphical abstract Image 1

SUBMITTER: Wu L

PROVIDER: S-EPMC9249825 | biostudies-literature |

REPOSITORIES: biostudies-literature

ACCESS DATA

Json Xml

Similar Datasets

An ultrasensitive molecularly imprinted polymer-based electrochemical sensor for the determination of SARS-CoV-2-RBD by using macroporous gold screen-printed electrode.

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

Reagentless Amperometric Pyruvate Biosensor Based on a Prussian Blue- and Enzyme Nanoparticle-Modified Screen-Printed Carbon Electrode.

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

Single-Drop Analysis of Epinephrine and Uric Acid on a Screen-Printed Carbon Electrode.

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.

| S-EPMC8392267 | biostudies-literature

Quantification of ethanol in plasma by electrochemical detection with an unmodified screen printed carbon electrode.

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

Direct Mercury Detection in Landfill Leachate Using a Novel AuNP-Biopolymer Carbon Screen-Printed Electrode Sensor.

Project description:A novel Au nanoparticle (AuNP)-biopolymer coated carbon screen-printed electrode (SPE) sensor was developed through the co-electrodeposition of Au and chitosan for mercury (Hg) ion detection. This new sensor showed successful Hg2+ detection in landfill leachate using square wave anodic stripping voltammetry (SWASV) with an optimized condition: a deposition potential of -0.6 V, deposition time of 200 s, amplitude of 25 mV, frequency of 60 Hz, and square wave step voltage of 4 mV. A noticeable peak was observed at +0.58 V associated with the stripping current of the Hg ion. The sensor exhibited a good sensitivity of ~0.09 μA/μg (~0.02 μA/nM) and a linear response over the concentration range of 10 to 100 ppb (50-500 nM). The limit of detection (LOD) was 1.69 ppb, which is significantly lower than the safety limit defined by the United States Environmental Protection Agency (USEPA). The sensor had an excellent selective response to Hg2+ in landfill leachate against other interfering cations (e.g., Zn2+, Pb2+, Cd2+, and Cu2+). Fifteen successive measurements with a stable peak current and a lower relative standard deviation (RSD = 5.1%) were recorded continuously using the AuNP-biopolymer-coated carbon SPE sensor, which showed excellent stability, sensitivity and reproducibility and consistent performance in detecting the Hg2+ ion. It also exhibited a good reliability and performance in measuring heavy metals in landfill leachate.

| S-EPMC8229311 | biostudies-literature

High-Sensitivity Dual-Probe Detection of Urinary miR-141 in Cancer Patients via a Modified Screen-Printed Carbon Electrode-Based Electrochemical Biosensor.

Project description:The screening and diagnosis of cancer are hallmarks of medicine in the aging population. Recently, microRNAs have shown potential for use as biomarkers, which could advance the field of diagnostics. The presence of miRNA-141 in the serum has been well described in several malignancies. However, the invasive approach used for sampling represents the major limitation for its practical application and, hence, its notable absence as a method for screening the general population. In light of this, we aimed to develop a high-sensitivity microRNA (miR) biosensor for application in the diagnosis of all miR-141-associated cancers, such as colorectal cancer (CRC) and breast cancer (BC). The novelty lies in our dual-probe design, which is reliant on the hybridization of the fluorescein isothiocyanate (FITC) targeting probe onto an existing sample of urinary miR-141 in the first step, followed by complementary binding with a biotinylated probe that has been coated on a modified screen-printed carbon electrode (SPCE). The hybridization of the probe and sensor produces signals via the catalytic reduction of H2O2 at HRP-modified SPCEs in the presence of H2O, which was measured by either cyclic voltammetry or chronoamperometry (CA) currents. In our study, the detection and expression of miR-141 in a cohort of colorectal cancer (n = 6) and breast cancer (n = 4) samples showed that its levels were significantly higher than in a healthy cohort (n = 9) (p < 0.004). Moreover, our miR sensor demonstrated high stability, reliability, and sensitivity (p < 0.0001). This work hopefully provides new information for the detection and monitoring of de novo and existing cancers.

| S-EPMC8125155 | biostudies-literature

Determination of Zinc, Cadmium, Lead, Copper and Silver Using a Carbon Paste Electrode and a Screen Printed Electrode Modified with Chromium(III) Oxide.

Project description:In this study, the preparation and electrochemical application of a chromium(III) oxide modified carbon paste electrode (Cr-CPE) and a screen printed electrode (SPE), made from the same material and optimized for the simple, cheap and sensitive simultaneous determination of zinc, cadmium, lead, copper and the detection of silver ions, is described. The limits of detection and quantification were 25 and 80 µg·L-1 for Zn(II), 3 and 10 µg·L-1 for Cd(II), 3 and 10 µg·L-1 for Pb(II), 3 and 10 µg·L-1 for Cu(II), and 3 and 10 µg·L-1 for Ag(I), respectively. Furthermore, this promising modification was transferred to the screen-printed electrode. The limits of detection for the simultaneous determination of zinc, cadmium, copper and lead on the screen printed electrodes were found to be 350 µg·L-1 for Zn(II), 25 µg·L-1 for Cd(II), 3 µg·L-1 for Pb(II) and 3 µg·L-1 for Cu(II). Practical usability for the simultaneous detection of these heavy metal ions by the Cr-CPE was also demonstrated in the analyses of wastewaters.

| S-EPMC5580028 | biostudies-literature

Magnetic beads combined with carbon black-based screen-printed electrodes for COVID-19: A reliable and miniaturized electrochemical immunosensor for SARS-CoV-2 detection in saliva.

Project description:The diffusion of novel SARS-CoV-2 coronavirus over the world generated COVID-19 pandemic event as reported by World Health Organization on March 2020. The huge issue is the high infectivity and the absence of vaccine and customised drugs allowing for hard management of this outbreak, thus a rapid and on site analysis is a need to contain the spread of COVID-19. Herein, we developed an electrochemical immunoassay for rapid and smart detection of SARS-CoV-2 coronavirus in saliva. The electrochemical assay was conceived for Spike (S) protein or Nucleocapsid (N) protein detection using magnetic beads as support of immunological chain and secondary antibody with alkaline phosphatase as immunological label. The enzymatic by-product 1-naphtol was detected using screen-printed electrodes modified with carbon black nanomaterial. The analytical features of the electrochemical immunoassay were evaluated using the standard solution of S and N protein in buffer solution and untreated saliva with a detection limit equal to 19 ng/mL and 8 ng/mL in untreated saliva, respectively for S and N protein. Its effectiveness was assessed using cultured virus in biosafety level 3 and in saliva clinical samples comparing the data using the nasopharyngeal swab specimens tested with Real-Time PCR. The agreement of the data, the low detection limit achieved, the rapid analysis (30 min), the miniaturization, and portability of the instrument combined with the easiness to use and no-invasive sampling, confer to this analytical tool high potentiality for market entry as the first highly sensitive electrochemical immunoassay for SARS-CoV-2 detection in untreated saliva.

| S-EPMC7833515 | biostudies-literature

Integrating PCR-free amplification and synergistic sensing for ultrasensitive and rapid CRISPR/Cas12a-based SARS-CoV-2 antigen detection.

Project description:Antigen detection provides particularly valuable information for medical diagnoses; however, the current detection methods are less sensitive and accurate than nucleic acid analysis. The combination of CRISPR/Cas12a and aptamers provides a new detection paradigm, but sensitive sensing and stable amplification in antigen detection remain challenging. Here, we present a PCR-free multiple trigger dsDNA tandem-based signal amplification strategy and a de novo designed dual aptamer synergistic sensing strategy. Integration of these two strategies endowed the CRISPR/Cas12a and aptamer-based method with ultra-sensitive, fast, and stable antigen detection. In a demonstration of this method, the limit of detection was at the single virus level (0.17 fM, approximately two copies/μL) in SARS-CoV-2 antigen nucleocapsid protein analysis of saliva or serum samples. The entire procedure required only 20 min. Given our system's simplicity and modular setup, we believe that it could be adapted reasonably easily for general applications in CRISPR/Cas12a-aptamer-based detection.

| S-EPMC8440162 | biostudies-literature

Fabrication of Nitrogen-Doped Reduced Graphene Oxide Modified Screen Printed Carbon Electrode (N-rGO/SPCE) as Hydrogen Peroxide Sensor.

Project description:In recent years, the electrochemical sensing approach has attracted electrochemists because of its excellent detection process, simplicity, high sensitivity, cost-effectiveness, and high selectivity. In this study, we prepared nitrogen doped reduced graphene oxide (N-rGO) and characterized it using various advanced techniques such as XRD, SEM, EDX, Raman, and XPS. Furthermore, we modified the active surface of a screen printed carbon electrode (SPCE) via the drop-casting of N-rGO. This modified electrode (N-rGO/SPCE) exhibited an excellent detection limit (LOD) of 0.83 µM with a decent sensitivity of 4.34 µAµM-1cm-2 for the detection of hydrogen peroxide (H2O2). In addition, N-rGO/SPCE also showed excellent selectivity, repeatability, and stability for the sensing of H2O2. Real sample investigations were also carried out that showed decent recovery.

| S-EPMC9324769 | biostudies-literature

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

Tweets

OmicsDI Databases

PRIDE
PeptideAtlas
MassIVE
JPOST Repository
Physiome Model Repository

EGA
EVA
ENA
LINCS
PAXDB
Cell Collective

MetaboLights
Metabolomics Workbench
MetabolomeExpress
GNPS
BioModels
FAIRDOMHub

ArrayExpress
dbGaP
ExpressionAtlas
GEO
NODE

Information

Databases
Help
API
Contact us
Code on GitHub
Terms of use
Submit Data