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1.
Int J Mol Sci ; 23(21)2022 Oct 28.
Article in English | MEDLINE | ID: covidwho-2090210

ABSTRACT

The global pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) caused a threat to public health and a worldwide crisis. This raised the need for quick, effective, and sensitive detection tools to prevent the rapid transmission rate of the infection. Therefore, this study aimed to develop an electrochemical impedance spectroscopy (EIS)-based aptasensor employing an interdigitated gold electrode (IDE) to detect SARS-CoV-2 Spike (S) glycoprotein and viral particles. This allowed us to sensitively detect SARS-CoV-2 S glycoprotein with a limit of detection (LOD) of 0.4 pg/mL in a buffer solution and to obtain a linear increase for concentrations between 0.2 to 0.8 pg/mL with high specificity. The proposed aptasensor also showed a good sensitivity towards the heat-inactivated SARS-CoV-2 variants in a buffer solution, where the Delta, Wuhan, and Alpha variants were captured at a viral titer of 6.45 ± 0.16 × 103 TCID50/mL, 6.20 × 104 TCID50/mL, and 5.32 ± 0.13 × 102 TCID50/mL, respectively. Furthermore, the detection of SARS-CoV-2 performed in a spiked human nasal fluid provided an LOD of 6.45 ± 0.16 × 103 TCID50/mL for the Delta variant in a 50 µL sample and a detection time of less than 25 min. Atomic force microscopy images complemented the EIS results in this study, revealing that the surface roughness of the IDE after each modification step increased, which indicates that the target was successfully captured. This label-free EIS-based aptasensor has promising potential for the rapid detection of SARS-CoV-2 in complex clinical samples.


Subject(s)
Biosensing Techniques , COVID-19 , Humans , SARS-CoV-2/genetics , Dielectric Spectroscopy , Biosensing Techniques/methods , COVID-19/diagnosis , Limit of Detection , Gold/chemistry , Electrodes , Electrochemical Techniques/methods
2.
Opt Express ; 30(22): 40277-40291, 2022 Oct 24.
Article in English | MEDLINE | ID: covidwho-2089309

ABSTRACT

We conceptualized and numerically investigated a photonic crystal fiber (PCF)-based surface plasmon resonance (SPR) sensor for rapid detection and quantification of novel coronavirus. The plasmonic gold-based optical sensor permits three different ways to quantify the virus concentrations inside patient's body based on different ligand-analyte conjugate pairs. This photonic biosensor demonstrates viable detections of SARS-CoV-2 spike receptor-binding-domain (RBD), mutated viral single-stranded ribonucleic acid (RNA) and human monoclonal antibody immunoglobulin G (IgG). A marquise-shaped core is introduced to facilitate efficient light-tailoring. Analytes are dissolved in sterile phosphate buffered saline (PBS) and surfaced on the plasmonic metal layer for realizing detection. The 1-pyrene butyric acid n-hydroxy-succinimide ester is numerically used to immobilize the analytes on the sensing interface. Using the finite element method (FEM), the proposed sensor is studied critically and optimized for the refractive index (RI) range from 1.3348-1.3576, since the target analytes RIs fluctuate within this range depending on the severity of the viral infection. The polarization-dependent sensor exhibits dominant sensing attributes for x-polarized mode, where it shows the average wavelength sensitivities of 2,009 nm/RIU, 2,745 nm/RIU and 1,984 nm/RIU for analytes: spike RBD, extracted coronavirus RNA and antibody IgG, respectively. The corresponding median amplitude sensitivities are 135 RIU-1, 196 RIU-1 and 140 RIU-1, respectively. The maximum sensor resolution and figure of merit are found 2.53 × 10-5 RIU and 101 RIU-1, respectively for viral RNA detection. Also, a significant limit of detection (LOD) of 6.42 × 10-9 RIU2/nm is obtained. Considering modern bioassays, the proposed compact photonic sensor will be well-suited for rapid point-of-care COVID testing.


Subject(s)
Biosensing Techniques , COVID-19 , Humans , SARS-CoV-2 , Ligands , Butyric Acid , COVID-19 Testing , RNA, Viral , COVID-19/diagnosis , Gold/chemistry , Immunoglobulin G , Succinimides , Pyrenes , Antibodies, Monoclonal , Esters , Phosphates
3.
Analyst ; 147(22): 5028-5037, 2022 Nov 07.
Article in English | MEDLINE | ID: covidwho-2069895

ABSTRACT

The continued uncertainty of emerging infectious viral diseases has led to an extraordinary urgency to develop advanced molecular diagnostic tests that are faster, more reliable, simpler to use, and readily available than traditional methods. This study presents a system that can accurately and rapidly trace viral nucleic acids by employing flap endonuclease 1 (FEN1)-assisted specific DNA cleavage reactions and surface-enhanced Raman scattering (SERS)-based analysis. The designed Raman tag-labeled 5'- and 3'-flap provider DNA yielded structurally defined DNA substrates on magnetic nanoparticle surfaces when a target was present. The FEN1 enzyme subsequently processes the substrates formed via an invasive cleavage reaction, producing 5'-flap DNA products. Magnetic separation allows efficient purification of flap products from reaction mixtures. The isolated solution was directly applied onto high aspect-ratio plasmonic silver nanopillars serving as SERS-active substrates to induce amplified SERS signals. We verified the developed SERS-based sensing system using a synthetic target complementary to an avian influenza A (H9N2) virus gene and examined the detection performance of the system using complementary DNA (cDNA) derived from H9N2 viral RNA. As a result, we could detect a synthetic target with a detection limit of 41.1 fM with a single base-pair discrimination ability and achieved multiplexed detection capability for two targets. Using cDNA samples from H9N2 viruses, we observed a high concordance of R2 = 0.917 between the data obtained from SERS and the quantitative polymerase chain reaction. We anticipate that this enzyme-assisted SERS sensor may provide insights into the development of high-performance molecular diagnostic tools that can respond rapidly to viral pathogens.


Subject(s)
Influenza A Virus, H9N2 Subtype , Metal Nanoparticles , Nucleic Acids , Animals , Spectrum Analysis, Raman/methods , Gold/chemistry , Flap Endonucleases , DNA, Complementary , DNA/analysis , Metal Nanoparticles/chemistry
4.
Opt Express ; 30(12): 22233-22246, 2022 Jun 06.
Article in English | MEDLINE | ID: covidwho-2065093

ABSTRACT

We propose a measurement method for sensitive and label-free detections of virus-like particles (VLPs) using color images of nanoplasmonic sensing chips. The nanoplasmonic chip consists of 5×5 gold nanoslit arrays and the gold surface is modified with specific antibodies for spike protein. The resonant wavelength of the 430-nm-period gold nanoslit arrays underwater environment is about 570 nm which falls between the green and red bands of the color CCD. The captured VLPs by the specific antibodies shift the plasmonic resonance of the gold nanoslits. It results in an increased brightness of green pixels and decreased brightness of red pixels. The image contrast signals of (green - red) / (red + green) show good linearity with the surface particle density. The experimental tests show the image contrast method can detect 100-nm polystyrene particles with a surface density smaller than 2 particles/µm2. We demonstrate the application for direct detection of SARS-CoV-2 VLPs using a simple scanner platform. A detection limit smaller than 1 pg/mL with a detection time less than 30 minutes can be achieved.


Subject(s)
Biosensing Techniques , COVID-19 , Nanostructures , Antibodies , Biosensing Techniques/methods , Gold/chemistry , Humans , Nanostructures/chemistry , Polystyrenes , SARS-CoV-2 , Spike Glycoprotein, Coronavirus , Surface Plasmon Resonance/methods
5.
Int J Mol Sci ; 23(18)2022 Sep 12.
Article in English | MEDLINE | ID: covidwho-2039868

ABSTRACT

The development of a strategy to investigate interfacial phenomena at lipid membranes is practically useful because most essential biomolecular interactions occur at cell membranes. In this study, a colorimetric method based on cysteine-encapsulated liposomes was examined using gold nanoparticles as a probe to provide a platform to report an enzymatic activity at lipid membranes. The cysteine-encapsulated liposomes were prepared with varying ratios of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) and cholesterol through the hydration of lipid films and extrusions in the presence of cysteine. The size, composition, and stability of resulting liposomes were analyzed by scanning electron microscopy (SEM), dynamic light scattering (DLS), nuclear magnetic resonance (NMR) spectroscopy, and UV-vis spectrophotometry. The results showed that the increased cholesterol content improved the stability of liposomes, and the liposomes were formulated with 60 mol % cholesterol for the subsequent experiments. Triton X-100 was tested to disrupt the lipid membranes to release the encapsulated cysteine from the liposomes. Cysteine can induce the aggregation of gold nanoparticles accompanying a color change, and the colorimetric response of gold nanoparticles to the released cysteine was investigated in various media. Except in buffer solutions at around pH 5, the cysteine-encapsulated liposomes showed the color change of gold nanoparticles only after being incubated with Triton X-100. Finally, the cysteine-encapsulated liposomal platform was tested to report the enzymatic activity of phospholipase A2 that hydrolyzes phospholipids in the membrane. The hydrolysis of phospholipids triggered the release of cysteine from the liposomes, and the released cysteine was successfully detected by monitoring the distinct red-to-blue color change of gold nanoparticles. The presence of phospholipase A2 was also confirmed by the appearance of a peak around 690 nm in the UV-vis spectra, which is caused by the cysteine-induced aggregation of gold nanoparticles. The results demonstrated that the cysteine-encapsulated liposome has the potential to be used to investigate biological interactions occurring at lipid membranes.


Subject(s)
Liposomes , Metal Nanoparticles , Cholesterol , Cysteine , Dimyristoylphosphatidylcholine , Gold/chemistry , Liposomes/chemistry , Metal Nanoparticles/chemistry , Octoxynol , Phospholipases , Phospholipids , Phosphorylcholine
6.
Sci Rep ; 12(1): 1724, 2022 02 02.
Article in English | MEDLINE | ID: covidwho-1663979

ABSTRACT

This study introduces localized surface plasmon resonance (L-SPR) mediated heating filter membrane (HFM) for inactivating universal viral particles by using the photothermal effect of plasmonic metal nanoparticles (NPs). Plasmonic metal NPs were coated onto filter membrane via a conventional spray-coating method. The surface temperature of the HFM could be controlled to approximately 40-60 °C at room temperature, owing to the photothermal effect of the gold (Au) NPs coated on them, under irradiation by visible light-emitting diodes. Due to the photothermal effect of the HFMs, the virus titer of H1Npdm09 was reduced by > 99.9%, the full inactivation time being < 10 min, confirming the 50% tissue culture infective dose (TCID50) assay. Crystal violet staining showed that the infectious samples with photothermal inactivation lost their infectivity against Mardin-Darby Canine Kidney cells. Moreover, photothermal inactivation could also be applied to reduce the infectivity of SARS-CoV-2, showing reduction rate of 99%. We used quantitative reverse transcription polymerase chain reaction (qRT-PCR) techniques to confirm the existence of viral genes on the surface of the HFM. The results of the TCID50 assay, crystal violet staining method, and qRT-PCR showed that the effective and immediate reduction in viral infectivity possibly originated from the denaturation or deformation of membrane proteins and components. This study provides a new, simple, and effective method to inactivate viral infectivity, leading to its potential application in various fields of indoor air quality control and medical science.


Subject(s)
COVID-19/virology , Hot Temperature , Light , Metal Nanoparticles , Micropore Filters , SARS-CoV-2 , Surface Plasmon Resonance/methods , Virion , Virus Inactivation , Air Pollution, Indoor , Animals , Cells, Cultured , Dogs , Gold/chemistry , SARS-CoV-2/genetics , SARS-CoV-2/pathogenicity
7.
Biosens Bioelectron ; 217: 114714, 2022 Dec 01.
Article in English | MEDLINE | ID: covidwho-2031161

ABSTRACT

Timely and accurate detection of virus is crucial for preventing spread of disease and early treatment of the infected cases. Herein we design an integrated logic-operated three-dimensional DNA walker for colorimetric detection of viral RNA fragments, by taking SARS-CoV-2 as an example. The DNA walker is composed of small amounts of dually-blocked walking strands and large amounts of dual-stem-loop track strands on gold nanoparticles. The walking strand contains a swing arm domain and a DNAzyme domain blocked at both sides of catalytic core, while the track strand contains a substrate domain located at the peripheral larger loop. Only the presence of both ORF1ab and N RNA fragments can fully de-block the walking strand, which then continuously hybridizes with track strands and cleaves them by DNAzyme-catalyzed hydrolysis. As the cleavage of track strands from long-stranded, double stem-loop structure to short-stranded, linear sequence, the DNA walker shows much lowered stability due to decreased negative charge density and diminished steric repulsion, which then gets aggregated at high salt concentration, accompanied by a visible color change. The colorimetric DNA walker detects RNA fragments down to 1 nM, responds dual viral genes in a "AND" logic way, and shows high specificity to target sequence. It can further detect large nucleic acids containing ORF1ab and N sequences, and reach 200 copies/mL detection limit by coupling a simple upstream amplification of sample. The method may provide a convenient way for reliable detection of viral RNA.


Subject(s)
Biosensing Techniques , COVID-19 , DNA, Catalytic , Metal Nanoparticles , Biosensing Techniques/methods , Colorimetry/methods , DNA/chemistry , DNA, Catalytic/chemistry , Gold/chemistry , Humans , Limit of Detection , Logic , Metal Nanoparticles/chemistry , RNA, Viral/genetics , SARS-CoV-2
8.
Dalton Trans ; 51(38): 14686-14699, 2022 Oct 04.
Article in English | MEDLINE | ID: covidwho-2028738

ABSTRACT

We report the controlled growth of biologically active compounds: gold nanoparticles (AuNPs) in various shapes, including their green synthesis, characterization, and studies of their applications towards biological, degradation and recycling. Using spectroscopic methods, studies on responsive binding mechanisms of AuNPs with biopolymers herring sperm deoxyribonucleic acid (hsDNA), bovine serum albumin (BSA), dyes degradation study, and exquisitely gold separation studies/recovery from nanowaste, COVID-19 testing kits, and pregnancy testing kits are discussed. The sensing ability of the AuNPs with biopolymers was investigated via various analytical techniques. The rate of degradation of various dyes in the presence and absence of AuNPs was studied by deploying stirring, IR, solar, and UV-Vis methods. AuNPs were found to be the most active cytotoxic agent against human breast cancer cell lines such as MCF-7 and MDAMB-468. Furthermore, an economical process for the recovery of gold traces from nanowaste, COVID-19 detection kits, and pregnancy testing kits was developed using inexpensive and eco-friendly α-cyclodextrin sugar. This method was found to be easy and safest in comparison with the universally accepted cyanidation process. In the future, small gold jewelry makers and related industries would benefit from the proposed gold-recycling process and it might contribute to their socio-economic growth. The methodologies proposed are also beneficial for trace-level forensic investigation.


Subject(s)
COVID-19 , Metal Nanoparticles , alpha-Cyclodextrins , COVID-19/diagnosis , COVID-19 Testing , Coloring Agents , Cytotoxins , DNA , Gold/chemistry , Humans , Male , Metal Nanoparticles/chemistry , Semen , Serum Albumin, Bovine/chemistry , Sugars
9.
ACS Nano ; 16(9): 14239-14253, 2022 Sep 27.
Article in English | MEDLINE | ID: covidwho-1991501

ABSTRACT

Limitations of the recognition elements in terms of synthesis, cost, availability, and stability have impaired the translation of biosensors into practical use. Inspired by nature to mimic the molecular recognition of the anti-SARS-CoV-2 S protein antibody (AbS) by the S protein binding site, we synthesized the peptide sequence of Asn-Asn-Ala-Thr-Asn-COOH (abbreviated as PEP2003) to create COVID-19 screening label-free (LF) biosensors based on a carbon electrode, gold nanoparticles (AuNPs), and electrochemical impedance spectroscopy. The PEP2003 is easily obtained by chemical synthesis, and it can be adsorbed on electrodes while maintaining its ability for AbS recognition, further leading to a sensitivity 3.4-fold higher than the full-length S protein, which is in agreement with the increase in the target-to-receptor size ratio. Peptide-loaded LF devices based on noncovalent immobilization were developed by affording fast and simple analyses, along with a modular functionalization. From studies by molecular docking, the peptide-AbS binding was found to be driven by hydrogen bonds and hydrophobic interactions. Moreover, the peptide is not amenable to denaturation, thus addressing the trade-off between scalability, cost, and robustness. The biosensor preserves 95.1% of the initial signal for 20 days when stored dry at 4 °C. With the aid of two simple equations fitted by machine learning (ML), the method was able to make the COVID-19 screening of 39 biological samples into healthy and infected groups with 100.0% accuracy. By taking advantage of peptide-related merits combined with advances in surface chemistry and ML-aided accuracy, this platform is promising to bring COVID-19 biosensors into mainstream use toward straightforward, fast, and accurate analyses at the point of care, with social and economic impacts being achieved.


Subject(s)
Biosensing Techniques , COVID-19 , Metal Nanoparticles , Biosensing Techniques/methods , COVID-19/diagnosis , COVID-19 Testing , Carbon/chemistry , Electrochemical Techniques , Electrodes , Gold/chemistry , Humans , Metal Nanoparticles/chemistry , Molecular Docking Simulation , Peptides/chemistry
10.
Biosensors (Basel) ; 12(8)2022 Aug 03.
Article in English | MEDLINE | ID: covidwho-1969093

ABSTRACT

In this research, we assessed the applicability of electrochemical sensing techniques for detecting specific antibodies against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike proteins in the blood serum of patient samples following coronavirus disease 2019 (COVID-19). Herein, screen-printed carbon electrodes (SPCE) with electrodeposited gold nanostructures (AuNS) were modified with L-Cysteine for further covalent immobilization of recombinant SARS-CoV-2 spike proteins (rSpike). The affinity interactions of the rSpike protein with specific antibodies against this protein (anti-rSpike) were assessed using cyclic voltammetry (CV) and differential pulse voltammetry (DPV) methods. It was revealed that the SPCE electroactive surface area increased from 1.49 ± 0.02 cm2 to 1.82 ± 0.01 cm2 when AuNS were electrodeposited, and the value of the heterogeneous electron transfer rate constant (k0) changed from 6.30 × 10-5 to 14.56 × 10-5. The performance of the developed electrochemical immunosensor was evaluated by calculating the limit of detection and limit of quantification, giving values of 0.27 nM and 0.81 nM for CV and 0.14 nM and 0.42 nM for DPV. Furthermore, a specificity test was performed with a solution of antibodies against bovine serum albumin as the control aliquot, which was used to assess nonspecific binding, and this evaluation revealed that the developed rSpike-based sensor exhibits low nonspecific binding towards anti-rSpike antibodies.


Subject(s)
Biosensing Techniques , COVID-19 , Nanostructures , Antibodies , Biosensing Techniques/methods , COVID-19/diagnosis , Carbon/chemistry , Electrochemical Techniques/methods , Electrodes , Gold/chemistry , Humans , Immunoassay/methods , Limit of Detection , SARS-CoV-2 , Spike Glycoprotein, Coronavirus
11.
Sci Rep ; 12(1): 8977, 2022 05 28.
Article in English | MEDLINE | ID: covidwho-1947469

ABSTRACT

Biosensors, especially those with a SERS readout, are required for an early and precise healthcare diagnosis. Unreproducible SERS platforms hamper clinical SERS. Here we report a synthetic procedure to obtain stabile, reproducible and robust highly-SERS performing nanocomposites for labelling. We controlled the NPs agglomeration and codification which resulted in an increased number of hot spots, thus exhibiting reproducible and superior Raman enhancement. We studied fundamental aspects affecting the plasmonic thiol bond resulting in pH exhibiting a determining role. We validated their biosensing performance by designing a SERS-based detection assay model for SARS-CoV-2. The limit of detection of our assay detecting the spike RBD was below 10 ng/mL.


Subject(s)
COVID-19 , Metal Nanoparticles , Gold/chemistry , Humans , Metal Nanoparticles/chemistry , SARS-CoV-2 , Spectrum Analysis, Raman/methods
12.
Biosens Bioelectron ; 213: 114476, 2022 Oct 01.
Article in English | MEDLINE | ID: covidwho-1944329

ABSTRACT

Coronavirus disease 2019 (COVID-19) has caused significant global morbidity and mortality. The serology test that detects antibodies against the disease causative agent, the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has often neglected value in supporting immunization policies and therapeutic decision-making. The ELISA-based antibody test is time-consuming and bulky. This work described a gold micro-interdigitated electrodes (IDE) biosensor for COVID antibody detection based on Electrochemical Impedance Spectroscopy (EIS) responses. The IDE architecture allows easy surface modification with the viral structure protein, Spike (S) protein, in the gap of the electrode digits to selectively capture anti-S antibodies in buffer solutions or human sera. Two strategies were employed to resolve the low sensitivity issue of non-faradic impedimetric sensors and the sensor fouling phenomenon when using the serum. One uses secondary antibody-gold nanoparticle (AuNP) conjugates to further distinguish anti-S antibodies from the non-specific binding and obtain a more significant impedance change. The second strategy consists of increasing the concentration of target antibodies in the gap of IDEs by inducing an AC electrokinetic effect such as dielectrophoresis (DEP). AuNP and DEP methods reached a limit of detection of 200 ng/mL and 2 µg/mL, respectively using purified antibodies in buffer, while the DEP method achieved a faster testing time of only 30 min. Both strategies could qualitatively distinguish COVID-19 antibody-positive and -negative sera. Our work, especially the impedimetric detection of COVID-19 antibodies under the assistance of the DEP force presents a promising path toward rapid, point-of-care solutions for COVID-19 serology tests.


Subject(s)
Biosensing Techniques , COVID-19 , Metal Nanoparticles , Biosensing Techniques/methods , COVID-19/diagnosis , Electrodes , Gold/chemistry , Humans , Metal Nanoparticles/chemistry , SARS-CoV-2
13.
Biosens Bioelectron ; 213: 114436, 2022 Oct 01.
Article in English | MEDLINE | ID: covidwho-1944325

ABSTRACT

The emergence of the COVID-19 epidemic has affected the lives of hundreds of millions of people globally. There is no doubt that the development of fast and sensitive detection methods is crucial while the worldwide effective vaccination programs are miles away from actualization. In this study, we have reported an electrochemical N protein aptamer sensor with complementary oligonucleotide as probe for the specific detection of COVID-19. The electrochemical aptasensor was prepared by fixing the double-stranded DNA hybrid obtained by the hybridization of N protein aptamer and its Fc-labeled complementary strand on the surface of a gold electrode. After incubation with the target, the aptamer dissociated from the labeled complementary DNA oligonucleotide hybrid to preferentially bind with N protein in the solution. The concentration of N protein was measured by detecting the changes in electrochemical current signals induced by the conformational transformation of the complementary DNA oligonucleotide left on the electrode surface. The sensor had a linear relationship between the logarithm of the N protein concentration from 10 fM to 100 nM (ΔIp = 0.098 log CN protein/fM - 0.08433, R2 = 0.99), and the detection limitation was 1 fM (S/N = 3). The electrochemical aptamer sensor was applied to test the spiked concentrations of throat swabs and blood samples from three volunteers, and the obtained results proved that the sensor has great potentials for the early detection of COVID-19 in patients.


Subject(s)
Aptamers, Nucleotide , Biosensing Techniques , COVID-19 , Aptamers, Nucleotide/chemistry , Biosensing Techniques/methods , COVID-19/diagnosis , DNA, Complementary , Electrochemical Techniques/methods , Electrodes , Gold/chemistry , Humans , Limit of Detection , Protein Binding
14.
Biosens Bioelectron ; 215: 114556, 2022 Nov 01.
Article in English | MEDLINE | ID: covidwho-1936098

ABSTRACT

Herein, an aptasensor was designed to detect the receptor-binding domain of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2-RBD) based on the encapsulation of the methylene blue (MB) inside the mesoporous silica film (MPSF), and an aptamer as an electrochemical probe, a porous matrix, and a bio-gatekeeper, respectively. The signal analysis of the proposed aptasensor indicated that the surface coverage of the encapsulated MB inside the MPSF (MB@MPSF) was 1.9 nmol/cm2. Aptamers were capped the MB@MPSF, avoiding the release of MB into the solution via the electrostatic attraction between the positively charged amino groups of the MPSF and negatively charged phosphate groups of the aptamers. Therefore, the electrochemical signal of the encapsulated MB in the absence of the SARS-CoV-2-RBD was high. In the presence of SARS-CoV-2-RBD, the aptamers that had a high affinity to the SARS-CoV-2-RBD molecules were removed from the electrode surface to interact with SARS-CoV-2-RBD. It gave rise to the release of the MB from the MPSF to the solution and washed away on the electrode surface. Therefore, the electrochemical signal of the aptasensor decreased. The electrochemical signal was recorded with a square wave voltammetry technical in the range of 0.5-250 ng/mL of SARS-CoV-2-RBD in a saliva sample. The limit of detection was found to be 0.36 ng/mL. Furthermore, the selectivity factor values of the proposed aptasensor to 32 ng/mL SARS-CoV-2-RBD in the presence of C-reactive protein, hemagglutinin, and neuraminidase of influenza A virus were 35.9, 11.7, and 17.37, respectively, indicating the high selectivity of the proposed aptasensor.


Subject(s)
Aptamers, Nucleotide , Biosensing Techniques , COVID-19 , Graphite , Aptamers, Nucleotide/chemistry , COVID-19/diagnosis , Electrochemical Techniques , Electrodes , Gold/chemistry , Graphite/chemistry , Humans , Lasers , Limit of Detection , Methylene Blue/chemistry , SARS-CoV-2 , Silicon Dioxide
15.
Mikrochim Acta ; 189(6): 226, 2022 05 20.
Article in English | MEDLINE | ID: covidwho-1941758

ABSTRACT

A promising immunosensing strategy in diagnosing SARS-CoV-2 is proposed using a 10-µm gap-sized gold interdigitated electrode (AuIDE) to target the surface spike protein (SP). The microelectrode surface was modified by (3-glycidyloxypropyl) trimethoxysilane to enforce the epoxy matrix, which facilitates the immobilization of the anti-SP antibody. The immunosensing performance was evaluated by integrating a nanosized (~ 10 nm) diamond-complexed SP as a target. The proposed immunoassay was quantitatively evaluated through electrochemical impedance spectroscopy (EIS) with the swept frequency from 0.1 to 1 MHz using a 100 mVRMS AC voltage supply. The immunoassay performed without diamond integration showed low sensitivity, with the lowest SP concentration measured at 1 pM at a determination coefficient of R2 = 0.9681. In contrast, the nanodiamond-conjugated SP on the immunosensor showed excellent sensitivity with a determination coefficient of R2 = 0.986. SP detection with a nanodiamond-conjugated target on AuIDE reached the low limit of detection at 189 fM in a linear detection range from 250 to 8000 fM. The specificity of the developed immunosensor was evaluated by interacting influenza-hemagglutinin and SARS-CoV-2-nucleocapsid protein with anti-SP. In addition, the authentic interaction of SP and anti-SP was validated by enzyme-linked immunosorbent assay.


Subject(s)
Biosensing Techniques , COVID-19 , Nanodiamonds , Biosensing Techniques/methods , COVID-19/diagnosis , Electric Impedance , Gold/chemistry , Humans , Immunoassay/methods , Microelectrodes , SARS-CoV-2 , Spike Glycoprotein, Coronavirus
16.
Environ Sci Pollut Res Int ; 29(58): 87764-87774, 2022 Dec.
Article in English | MEDLINE | ID: covidwho-1930520

ABSTRACT

Pharmaceutical products are some of the most serious emergent pollutants in the environment, especially nowadays of the COVID-19 pandemic. In this study, nanogold-composite was prepared, and its catalytic activity for paracetamol degradation was investigated. Moreover, for the first time, recycled waste diatomite earth (WDE) from beer filtration was used for reproducible gold nanoparticle (Au NPs) preparation. We studied Au NPs by various psychical-chemical and analytical methods. Transmission and scanning electron microscopy were used for nanogold-composite morphology, size and shape characterization. Total element concentrations were determined using inductively coupled plasma mass and X-ray fluorescence spectrometry. X-ray powder diffraction analysis was used for crystal structure characterization of samples. Fourier transform infrared spectrometer was used to study the chemical changes before and after Au NP formation. The results revealed that the WDE served as both a reducing and a stabilizing agent for crystalline spherical 30 nm Au NPs as well as acting as a direct support matrix. The kinetics of paracetamol degradation was studied by high-performance liquid chromatography with a photodiode array detector. The conversion of paracetamol was 62% and 67% after 72 h in the absence or presence of light irradiation, respectively, with 0.0126 h-1 and 0.0148 h-1 reaction rate constants. The presented study demonstrates the successful use of waste material from the food industry for nanogold-composite preparation and its application as a promising catalyst in paracetamol removal.


Subject(s)
COVID-19 , Metal Nanoparticles , Humans , Gold/chemistry , Metal Nanoparticles/chemistry , Acetaminophen/chemistry , Pandemics
17.
Wiley Interdiscip Rev Nanomed Nanobiotechnol ; 14(5): e1817, 2022 09.
Article in English | MEDLINE | ID: covidwho-1913900

ABSTRACT

Gold and silver nanoparticles (NPs) are widely used in the biomedical research both in the therapeutic and the sensing/diagnostics fronts. Both metals share some common optical properties with surface plasmon resonance being the most widely exploited property in therapeutics and diagnostics. Au NPs exhibit excellent light-to-heat conversion efficiencies and hence have found applications primarily in precision oncology, while Ag NPs have excellent antibacterial properties which can be harnessed in biomaterials' design. Both metals constitute excellent biosensing platforms owing to their plasmonic properties and are now routinely used in various optical platforms. The utilization of Au and Ag NPs in the COVID-19 pandemic was rapidly expanded mostly in biosensing and point-of-care platforms and to some extent in therapeutics. In this review article, the main physicochemical properties of Au and Ag NPs are discussed with selective examples from the recent literature. This article is categorized under: Therapeutic Approaches and Drug Discovery > Nanomedicine for Oncologic Disease Diagnostic Tools > In Vitro Nanoparticle-Based Sensing Nanotechnology Approaches to Biology > Nanoscale Systems in Biology.


Subject(s)
COVID-19 , Metal Nanoparticles , Neoplasms , Gold/chemistry , Humans , Metal Nanoparticles/chemistry , Neoplasms/drug therapy , Pandemics , Precision Medicine , Silver
18.
Anal Bioanal Chem ; 414(18): 5507-5517, 2022 Jul.
Article in English | MEDLINE | ID: covidwho-1906016

ABSTRACT

This paper reports the development of a low-cost (< US$ 0.03 per device) immunosensor based on gold-modified screen-printed carbon electrodes (SPCEs). As a proof of concept, the immunosensor was tested for a fast and sensitive determination of S proteins from both SARS-CoV and SARS-CoV-2, by a single disposable device. Gold nanoparticles were electrochemically deposited via direct reduction of gold ions on the electrode using amperometry. Capture antibodies from spike (S) protein were covalently immobilized on carboxylic groups of self-assembled monolayers (SAM) of mercaptoacetic acid (MAA) attached to the gold nanoparticles. Label-free detection of S proteins from both SARS-CoV and SARS-CoV-2 was performed with electrochemical impedance spectroscopy (EIS). The immunosensor fabricated with 9 s gold deposition had a high performance in terms of selectivity, sensitivity, and low limit of detection (LOD) (3.16 pmol L-1), thus permitting the direct determination of the target proteins in spiked saliva samples. The complete analysis can be carried out within 35 min using a simple one-step assay protocol with small sample volumes (10 µL). With such features, the immunoplatform presented here can be deployed for mass testing in point-of-care settings.


Subject(s)
Biosensing Techniques , COVID-19 , Metal Nanoparticles , Nanostructures , SARS Virus , Biosensing Techniques/methods , COVID-19/diagnosis , Electrochemical Techniques/methods , Electrodes , Gold/chemistry , Humans , Immunoassay/methods , Limit of Detection , Metal Nanoparticles/chemistry , SARS-CoV-2
19.
SLAS Discov ; 27(6): 331-336, 2022 09.
Article in English | MEDLINE | ID: covidwho-1878377

ABSTRACT

Current methods for the screening of viral infections in clinical settings, such as reverse transcription polymerase chain reaction (RT-qPCR) and enzyme-linked immunosorbent assay (ELISA), are expensive, time-consuming, require trained personnel and sophisticated instruments. Therefore, novel sensors that can save time and cost are required specially in remote areas and developing countries that may lack the advanced scientific infrastructure for this task. In this work, we present a sensitive, and highly specific biosensing approach for the detection of harmful viruses that have cysteine residues within the structure of their cell surface proteins. We utilized new method for the rapid screening of SARS-CoV-2 virus in biological fluids through its S1 protein by surface enhanced Raman spectroscopy (SERS). The protein is captured from aqueous solutions and biological specimens using a target-specific extractor substrate. The structure of the purified protein is then modified to convert it into a bio-thiol by breaking the disulfide bonds and freeing up the sulfhydryl (SH) groups of the cysteine residues. The formed biothiol chemisorbs favourably onto a highly sensitive plasmonic sensor and probed by a handheld Raman device in few seconds. The new method was used to screen the S1 protein in aqueous medium, spiked human blood plasma, mucus, and saliva samples down to 150 fg/L. The label-free SERS biosensing method has strong potential for the fingerprint identification many viruses (e.g. the human immunodeficiency virus, the human polyomavirus, the human papilloma virus, the adeno associated viruses, the enteroviruses) through the cysteine residues of their capsid proteins. The new method can be applied at points of care (POC) in remote areas and developing countries lacking sophisticated scientific infrastructure.


Subject(s)
COVID-19 , SARS-CoV-2 , COVID-19/diagnosis , Cysteine , Gold/chemistry , Humans , Limit of Detection , Membrane Proteins
20.
ACS Appl Bio Mater ; 5(5): 2421-2430, 2022 05 16.
Article in English | MEDLINE | ID: covidwho-1829968

ABSTRACT

In this work, we report a facile synthesis of graphene oxide-gold (GO-Au) nanocomposites by electrodeposition. The fabricated electrochemical immunosensors are utilized for the dual detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antigen and SARS-CoV-2 antibody. The GO-Au nanocomposites has been characterized by UV-vis spectroscopy, X-ray diffraction (XRD), transmission electron microscopy (TEM), cyclic voltammetry (CV), differential pulse voltammetry (DPV), and electrochemical impedance spectroscopy (EIS) for its biosensing properties. The linear detection range of the SARS-CoV-2 antigen immunosensor is 10.0 ag mL-1 to 50.0 ng mL-1, whereas that for the antibody immunosensor ranges from 1.0 fg mL-1 to 1.0 ng mL-1. The calculated limit of detection (LOD) of the SARS-CoV-2 antigen immunosensor is 3.99 ag mL-1, and that for SARS-CoV-2 antibody immunosensor is 1.0 fg mL-1 with high sensitivity. The validation of the immunosensor has also been carried out on patient serum and patient swab samples from COVID-19 patients. The results suggest successful utilization of the immunosensors with a very low detection limit enabling its use in clinical samples. Further work is needed for the standardization of the results and translation in screen-printed electrodes for use in portable commercial applications.


Subject(s)
Biosensing Techniques , COVID-19 , Metal Nanoparticles , Nanocomposites , Antibodies , Biosensing Techniques/methods , COVID-19/diagnosis , Gold/chemistry , Graphite , Humans , Immunoassay/methods , Metal Nanoparticles/chemistry , Nanocomposites/chemistry , SARS-CoV-2
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