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1.
ACS Appl Mater Interfaces ; 14(31): 35299-35308, 2022 Aug 10.
Article in English | MEDLINE | ID: covidwho-1960239

ABSTRACT

Loop-mediated isothermal amplification (LAMP) has received considerable attention for decentralized (point-of-care and on-site) nucleic acid testing in view of its simple temperature control (60-65 °C) and short assay time (15-60 min). There remains a challenge in its wide adoption and acceptance due to the limitations of the existing amplification result reporter probes, e.g., photobleaching of organic fluorophore and reduced sensitivity of the pH-sensitive colorimetric dye. Herein, we demonstrate CdSeS/ZnS quantum dots (semiconductor fluorescent nanocrystals with superior photostability than organic fluorophore) with surface modification of cysteamine (amine-QDs) as a new reporter probe for LAMP that enabled single-copy sensitivity (limit of detection of 83 zM; 20 µL reaction volume). For a negative LAMP sample (absence of target sequence), positively charged amine-QDs remained dispersed due to interparticle electrostatic repulsion. While for a positive LAMP sample (presence of target sequence), amine-QDs became precipitated. The characterization data showed that amine-QDs were embedded in magnesium pyrophosphate crystals (generated during positive LAMP), thus leading to their coprecipitation. This amine-QD-based one-step LAMP assay advances the field of QD-based nucleic acid amplification assays in two aspects: (1) compatibility─one-step amplification and detection (versus separation of amplification and detection steps); and (2) universality─the same amine-QDs for different target sequences (versus different oligonucleotide-modified QDs for different target sequences).


Subject(s)
Nucleic Acids , Quantum Dots , Amines , Molecular Diagnostic Techniques , Nucleic Acid Amplification Techniques , Sensitivity and Specificity
2.
Anal Methods ; 14(26): 2631-2641, 2022 07 07.
Article in English | MEDLINE | ID: covidwho-1900675

ABSTRACT

In this work, a simple, low-cost and easy-to-handle analytical procedure based on carbon quantum dots (CQDs) is proposed to check commercially available formulated microbicides that are used to mitigate the transmission of viruses, such as SARS-COV-2, or bacterial diseases. For this purpose, CQDs were synthesized via pyrolysis using citric acid and ethylenediamine as precursors to produce an intense fluorescence that is used to measure the concentration of hypochlorite, an important biocidal agent present in sanitizing mats, by quenching mechanisms. The characterization of the CQDs was performed using IR spectrophotometry, UV-Vis spectrophotometry, spectrofluorometry, thermogravimetric analysis, scanning electron microscopy, dynamic light scattering, X-ray diffraction, energy-dispersive spectroscopy, and zeta potential measurements. For analytical purposes, fluorescence was measured in a UV chamber irradiated using an LED with the maximum emission at 350 nm. A smartphone was coupled to the UV chamber to measure the fluorescence quenching due to the presence of hypochlorite, and further the digital images were decomposed by RGB data using free software. Tests of pH, CQD concentration and stability of the fluorescence emitted were performed. The stability study of the fluorescence emitted by the CQD solution showed a relative standard deviation lower than 5.0%. The fluorescence digital image-based (FDIB) method resulted in a linear range from 17.44 µmol L-1 to 90.0 µmol L-1 with an LOD of 3.30 µmol L-1 for the determination of hypochlorite using a microplate made of PLA (polylactic acid) customized using a 3D printer. Furthermore, the hypochlorite concentration was tested in situ for its compliance in a sanitizing mat, in a real use situation (daily, a group of four people, each one kept their feet on the mat for 30 s). After 2.5 h, the monitored concentration of hypochlorite was 0.04953% (w/v) or 7.63 mmol L-1, and therefore, it was inefficient to act as a sanitizing agent. Thus, for the first time in the literature, an FDIB method with CQDs is used to verify in situ microbicide practices with a fast and low-cost analytical procedure.


Subject(s)
COVID-19 , Quantum Dots , Carbon/chemistry , Carbon/pharmacology , Humans , Hypochlorous Acid , Quantum Dots/chemistry , SARS-CoV-2
3.
Lett Appl Microbiol ; 74(6): 1001-1007, 2022 Jun.
Article in English | MEDLINE | ID: covidwho-1891648

ABSTRACT

African swine fever (ASF), a highly contagious and lethal disease, poses a tremendous threat and burden to the swine industry worldwide. Lack of available vaccines or treatments leaves rapid diagnosis as the key tool to control the disease. Quantum dots (QDs) are unique fluorescent semiconductor nanoparticles, highly versatile for biological applications. In this study, we developed a quantum dots-based fluorescent immunochromatographic assay (QDs-FICA) using CD2v as the diagnosis antigen to detect ASFV antibodies. The titre of the test strip was 1 : 5·12 × 105 . In addition, the strip was highly specific to anti-ASFV serum and had no cross-reaction with CSFV, PPV, PRRSV, PCV-2, PRV and FMDV. Moreover, a comparative test of 71 clinical samples showed that the coincidence rate was 85·92% between the test strip and the commercial ELISA kit (coated with p30, p62 and p72). The QDs-FICA can be used to detect ASFV antibodies, which is meaningful for the surveillance, control and purification of ASF.


Subject(s)
African Swine Fever Virus , African Swine Fever , Quantum Dots , African Swine Fever/diagnosis , African Swine Fever/prevention & control , Animals , Diagnosis, Differential , Immunoassay , Swine
4.
Int J Mol Sci ; 23(11)2022 Jun 02.
Article in English | MEDLINE | ID: covidwho-1884205

ABSTRACT

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is the pathogenic agent leading to COVID-19. Due to high speed of transmission and mutation rates, universal diagnosis and appropriate prevention are still urgently needed. The nucleocapsid protein of SARS-CoV-2 is considered more conserved than spike proteins and is abundant during the virus' life cycle, making it suitable for diagnostic applications. Here, we designed and developed a fluorescent immunochromatography assay (FICA) for the rapid detection of SARS-CoV-2-specific antibodies using ZnCdSe/ZnS QDs-conjugated nucleocapsid (N) proteins as probes. The nucleocapsid protein was expressed in E.coli and purified via Ni-NTA affinity chromatography with considerable concentration (0.762 mg/mL) and a purity of more than 90%, which could bind to specific antibodies and the complex could be captured by Staphylococcal protein A (SPA) with fluorescence displayed. After the optimization of coupling and detecting conditions, the limit of detection was determined to be 1:1.024 × 105 with an IgG concentration of 48.84 ng/mL with good specificity shown to antibodies against other zoonotic coronaviruses and respiratory infection-related viruses (n = 5). The universal fluorescent immunochromatography assay simplified operation processes in one step, which could be used for the point of care detection of SARS-CoV-2-specific antibodies. Moreover, it was also considered as an efficient tool for the serological screening of potential susceptible animals and for monitoring the expansion of virus host ranges.


Subject(s)
COVID-19 , Quantum Dots , Animals , Antibodies, Viral , COVID-19/diagnosis , Chromatography, Affinity , Nucleocapsid Proteins , SARS-CoV-2 , Sensitivity and Specificity
5.
Viruses ; 14(5)2022 05 12.
Article in English | MEDLINE | ID: covidwho-1869809

ABSTRACT

A new antibody diagnostic assay with more rapid and robust properties is demanded to quantitatively evaluate anti-severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) immunity in a large population. Here, we developed a nanometer-scale fluorescent biosensor system consisting of CdSe-ZnS quantum dots (QDs) coupled with the highly sensitive B-cell epitopes of SARS-CoV-2 that could remarkably identify the corresponding antibody with a detection limit of 100 pM. Intriguingly, we found that fluorescence quenching of QDs was stimulated more obviously when coupled with peptides than the corresponding proteins, indicating that the energy transfer between QDs and peptides was more effective. Compared to the traditional enzyme-linked immunosorbent assay (ELISA), the B-cell-epitope-based QD-biosensor could robustly distinguish coronavirus disease 2019 (COVID-19) antibody-positive patients from uninfected individuals with a higher sensitivity (92.3-98.1% positive rates by QD-biosensor vs. 78.3-83.1% positive rates by ELISAs in 207 COVID-19 patients' sera) in a more rapid (5 min) and labor-saving manner. Taken together, the 'QD-peptides' biosensor provided a novel real-time, quantitative, and high-throughput method for clinical diagnosis and home-use tests.


Subject(s)
Biosensing Techniques , COVID-19 , Quantum Dots , Antibodies , COVID-19/diagnosis , Epitopes, B-Lymphocyte , Humans , Peptides , SARS-CoV-2
6.
Biosens Bioelectron ; 207: 114209, 2022 Jul 01.
Article in English | MEDLINE | ID: covidwho-1838599

ABSTRACT

The sudden increase of the COVID-19 outbreak and its continued growth with mutations in various forms has created a global health crisis as well as devastating social and economic effects over the past two years. In this study, a screen-printed carbon electrode reinforced with boron nitride quantum dots/flower-like gold nanostructures (BNQDs/FGNs/SPCE) and functionalized by highly specific antisense DNA oligonucleotide presents an alternative and promising solution for targeting SARS-CoV-2 RNA without nucleic acid amplification. The platform was tested on 120 SARS-CoV-2 RNA isolated from real clinical samples (60 positive and 60 negative confirmed by conventional RT-PCR method). Based on obtained quantitative results and statistical analysis (box-diagram, cutoff value, receiver operating characteristic curve, and t-test), the biosensor revealed a significant difference between the two positive and negative groups with 100% sensitivity and 100% specificity. To evaluate the quantitation capacity and detection limit of the biosensor for clinical trials, the detection performance of the biosensor for continuously diluted RNA isolated from SARS-CoV-2-confirmed patients was compared to those obtained by RT-PCR, demonstrating that the detection limit of the biosensor is lower than or comparable to that of RT-PCR. The ssDNA/BNQDs/FGNs/SPCE showed negligible cross-reactivity with RNA fragments isolated from Influenza A (IAV) clinical samples and also remained stable for up to 14 days. In conclusion, the fabricated biosensor may serve as a promising tool for point-of-care applications.


Subject(s)
Biosensing Techniques , COVID-19 , Nanostructures , Quantum Dots , Biosensing Techniques/methods , Boron Compounds , COVID-19/diagnosis , Gold , Humans , RNA, Viral/genetics , SARS-CoV-2/genetics , Sensitivity and Specificity
7.
J Vis Exp ; (182)2022 04 21.
Article in English | MEDLINE | ID: covidwho-1834949

ABSTRACT

The development of new technologies for cellular fluorescence microscopy has facilitated high-throughput screening methods for drug discovery. Quantum dots are fluorescent nanoparticles with excellent photophysical properties imbued with bright and stable photoluminescence as well as narrow emission bands. Quantum dots are spherical in shape, and with the proper modification of the surface chemistry, can be used to conjugate biomolecules for cellular applications. These optical properties, combined with the ability to functionalize them with biomolecules, make them an excellent tool for investigating receptor-ligand interactions and cellular trafficking. Here, we present a method that uses quantum dots to track the binding and endocytosis of SARS-CoV-2 spike protein. This protocol can be used as a guide for experimentalists looking to utilize quantum dots to study protein-protein interactions and trafficking in the context of cellular physiology.


Subject(s)
Endocytosis , Quantum Dots , Spike Glycoprotein, Coronavirus , HEK293 Cells , Humans , SARS-CoV-2 , Spike Glycoprotein, Coronavirus/analysis
8.
Int J Infect Dis ; 121: 58-65, 2022 Aug.
Article in English | MEDLINE | ID: covidwho-1804270

ABSTRACT

BACKGROUND: As several vaccines for SARS-CoV-2 have been developed, a large proportion of individuals have been vaccinated worldwide so far. The rapid and accurate immunoassays are urgently needed for detecting the specific virus-neutralizing antibody (NAb), which reflect the protective effect of the vaccines among different populations. METHODS: In this study, we designed a quantum dot lateral flow immunoassay strip (QD-LFIA) for smartphones for the detection of specific IgG or neutralizing antibodies in SARS-CoV-2 in human serum or whole blood samples. The recombinant receptor binding domain of the SARS-CoV-2 spike protein was used as the antigen to combine with NAb or angiotensin-converting enzyme 2. RESULTS: Among 81 patients who recovered from COVID-19 who were diagnosed using the nucleic acid test initially, 98.8% (80/81) were positive for IgG and 88.9% (72/81) were positive for NAb by QD-LFIA. Among 64 individuals inoculated with inactivated vaccines and six subunit vaccines, 90% (63/70) were positive for IgG and 82.9% (58/70) were positive for NAb by QD-LFIA, whereas no cross-reaction was found in 150 healthy blood donors, two patients with influenza B, and three patients with common cold. CONCLUSION: The established platform could achieve a rapid and accurate detection of NAb specific to SARS-CoV-2, which could be used for detecting the protective effect of the vaccines in areas of world that currently affected by the pandemic.


Subject(s)
COVID-19 , Quantum Dots , Antibodies, Neutralizing , Antibodies, Viral , COVID-19/diagnosis , COVID-19 Vaccines , Humans , Immunoassay , Immunoglobulin G , SARS-CoV-2 , Smartphone , Spike Glycoprotein, Coronavirus
9.
Int J Biol Macromol ; 206: 115-147, 2022 May 01.
Article in English | MEDLINE | ID: covidwho-1697104

ABSTRACT

Thanks to their unique attributes, such as good sensitivity, selectivity, high surface-to-volume ratio, and versatile optical and electronic properties, fluorescent-based bioprobes have been used to create highly sensitive nanobiosensors to detect various biological and chemical agents. These sensors are superior to other analytical instrumentation techniques like gas chromatography, high-performance liquid chromatography, and capillary electrophoresis for being biodegradable, eco-friendly, and more economical, operational, and cost-effective. Moreover, several reports have also highlighted their application in the early detection of biomarkers associated with drug-induced organ damage such as liver, kidney, or lungs. In the present work, we comprehensively overviewed the electrochemical sensors that employ nanomaterials (nanoparticles/colloids or quantum dots, carbon dots, or nanoscaled metal-organic frameworks, etc.) to detect a variety of biological macromolecules based on fluorescent emission spectra. In addition, the most important mechanisms and methods to sense amino acids, protein, peptides, enzymes, carbohydrates, neurotransmitters, nucleic acids, vitamins, ions, metals, and electrolytes, blood gases, drugs (i.e., anti-inflammatory agents and antibiotics), toxins, alkaloids, antioxidants, cancer biomarkers, urinary metabolites (i.e., urea, uric acid, and creatinine), and pathogenic microorganisms were outlined and compared in terms of their selectivity and sensitivity. Altogether, the small dimensions and capability of these nanosensors for sensitive, label-free, real-time sensing of chemical, biological, and pharmaceutical agents could be used in array-based screening and in-vitro or in-vivo diagnostics. Although fluorescent nanoprobes are widely applied in determining biological macromolecules, unfortunately, they present many challenges and limitations. Efforts must be made to minimize such limitations in utilizing such nanobiosensors with an emphasis on their commercial developments. We believe that the current review can foster the wider incorporation of nanomedicine and will be of particular interest to researchers working on fluorescence technology, material chemistry, coordination polymers, and related research areas.


Subject(s)
Biosensing Techniques , Nanoparticles , Nanostructures , Quantum Dots , Biosensing Techniques/methods , Carbon/chemistry , Coloring Agents
10.
Biosens Bioelectron ; 202: 113978, 2022 Apr 15.
Article in English | MEDLINE | ID: covidwho-1661800

ABSTRACT

The development of reliable, sensitive, and fast devices for the diagnosis of COVID-19 is of great importance in the pandemic of the new coronavirus. Here, we proposed a new principle of analysis based on a combination of reverse transcription and isothermal amplification of a fragment of the gene encoding the S protein of the SARS-CoV-2 and the CRISPR/Cas13a reaction for cleavage of the specific probe. As a result, the destroyed probe cannot be detected on an immunochromatographic strip using quantum fluorescent dots. Besides, the results can be obtained by an available and inexpensive portable device. By detecting SARS-CoV-2 negative (n = 25) and positive (n = 62) clinical samples including throat swabs, sputum and anal swabs, the assay showed good sensitivity and specificity of the method and could be completed within 1 h without complicated operation and expensive equipment. These superiorities showed its potential for fast point-of-care screening of SARS-CoV-2 during the outbreak, especially in remote and underdeveloped areas with limited equipment and resources.


Subject(s)
Biosensing Techniques , COVID-19 , Quantum Dots , Chromatography, Affinity , Clustered Regularly Interspaced Short Palindromic Repeats , Humans , Nucleic Acid Amplification Techniques/methods , RNA, Viral/genetics , SARS-CoV-2 , Sensitivity and Specificity
11.
Biosens Bioelectron ; 202: 113974, 2022 Apr 15.
Article in English | MEDLINE | ID: covidwho-1611633

ABSTRACT

Rapid and reliable detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibody can provide immunological evidence in addition to nucleic acid test for the early diagnosis and on-site screening of coronavirus disease 2019 (COVID-19). All-solid-state biosensor capable of rapid, quantitative SARS-CoV-2 antibody testing is still lacking. Herein, we propose an electronic labelling strategy of protein molecules and demonstrate SARS-CoV-2 protein biosensor employing colloidal quantum dots (CQDs)-modified electrode. The feature current peak corresponding to the specific binding reaction of SARS-CoV-2 antigen and antibody proteins was observed for the first time. The unique charging and discharging effect depending on the alternating voltage applied was ascribed to the quantum confinement, Coulomb blockade and quantum tunneling effects of quantum dots. CQDs-modified electrode could recognize the specific binding reaction between antigen and antibody and then transduce it into significant electrical current. In the case of serum specimens from COVID-19 patient samples, the all-solid-state protein biosensor provides quantitative analysis of SARS-CoV-2 antibody with correlation coefficient of 93.8% compared to enzyme-linked immunosorbent assay (ELISA) results. It discriminates patient and normal samples with accuracy of about 90%. The results could be read within 1 min by handheld testing system prototype. The sensitive and specific protein biosensor combines the advantages of rapidity, accuracy, and convenience, facilitating the implement of low-cost, high-throughput immunological diagnostic technique for clinical lab, point-of-care testing (POCT) as well as home-use test.


Subject(s)
Biosensing Techniques , COVID-19 , Quantum Dots , Biosensing Techniques/methods , Electrodes , Humans , SARS-CoV-2 , Sensitivity and Specificity
12.
Mikrochim Acta ; 188(10): 316, 2021 Sep 02.
Article in English | MEDLINE | ID: covidwho-1604245

ABSTRACT

A novel label-free surface plasmon resonance (SPR) aptasensor has been constructed for the detection of N-gene of SARS-CoV-2 by using thiol-modified niobium carbide MXene quantum dots (Nb2C-SH QDs) as the bioplatform for anchoring N-gene-targeted aptamer. In the presence of SARS-CoV-2 N-gene, the immobilized aptamer strands changed their conformation to specifically bind with N-gene. It thus increased the contact area or enlarged the distance between aptamer and the SPR chip, resulting in a change of the SPR signal irradiated by the laser (He-Ne) with the wavelength (λ) of 633 nm. Nb2C QDs were derived from Nb2C MXene nanosheets via a solvothermal method, followed by functionalization with octadecanethiol through a self-assembling method. Subsequently, the gold chip for SPR measurements was modified with Nb2C-SH QDs via covalent binding of the Au-S bond also by self-assembling interaction. Nb2C-SH QDs not only resulted in high bioaffinity toward aptamer but also enhanced the SPR response. Thus, the Nb2C-SH QD-based SPR aptasensor had low limit of detection (LOD) of 4.9 pg mL-1 toward N-gene within the concentration range 0.05 to 100 ng mL-1. The sensor also showed excellent selectivity in the presence of various respiratory viruses and proteins in human serum and high stability. Moreover, the Nb2C-SH QD-based SPR aptasensor displayed a vast practical application for the qualitative analysis of N-gene from different samples, including seawater, seafood, and human serum. Thus, this work can provide a deep insight into the construction of the aptasensor for detecting SARS-CoV-2 in complex environments. A novel label-free surface plasmon resonance aptasensor has been constructed to detect sensitively and selectively the N-gene of SARS-CoV-2 by using thiol-modified niobium carbide MXene quantum dots as the scaffold to anchor the N-gene-targeted aptamer.


Subject(s)
Aptamers, Nucleotide , COVID-19/diagnosis , Niobium/chemistry , Nucleocapsid/metabolism , Quantum Dots/chemistry , SARS-CoV-2/isolation & purification , Surface Plasmon Resonance/methods , COVID-19/virology , Humans , Limit of Detection
13.
Spectrochim Acta A Mol Biomol Spectrosc ; 269: 120702, 2022 Mar 15.
Article in English | MEDLINE | ID: covidwho-1559881

ABSTRACT

Urgent identification of COVID-19 in infected patients is highly important nowadays. Förster or fluorescence resonance energy transfer (FRET) is a powerful and sensitive method for nanosensing applications, and quantum dots are essential materials in FRET-based nanosensors. The QDs are conjugated to DNA or RNA and used in many applications. Therefore, in the present study, novel fluorescence DNA-conjugated CdTe/ZnS quantum dots nanoprobe designed for detection of Covid-19 after extracting their RNA from saliva of hesitant people. For achieving this purpose, the water-soluble CdTe/ZnS QDs-DNA prepared via replacing the thioglycolic acid (TGA) on the surface of QDs with capture DNA (thiolated DNA) throw a ligand-exchange method. Subsequently, by adding the different concentrations of complementary (target DNA) in a mixture of quencher DNA (BHQ2-labeled DNA) and the QDs-DNA conjugates at different conditions, sandwiched hybrids were formed. The results showed that the fluorescence intensity was decreased with increasing the concentration of target DNA (as a positive control). The linear equation and regression (Y = 40.302 X  + 1 and R2 = 0.98) were obtained by using the Stern-Volmer relationship. The Limit of detection (LOD) was determined 0.000823 µM. The achieved results well confirm the outcomes of the RT-PCR method in real samples.


Subject(s)
COVID-19 , Cadmium Compounds , Quantum Dots , DNA , Humans , SARS-CoV-2 , Sulfides , Tellurium , Zinc Compounds
14.
Expert Opin Drug Discov ; 17(3): 225-230, 2022 Mar.
Article in English | MEDLINE | ID: covidwho-1532354

ABSTRACT

INTRODUCTION: SARS-CoV-2 is a highly infectious and deadly coronavirus whose study requires the use of a biosafety level 3 (BSL-3) containment facility to investigate viral biology and pathogenesis, which limits the study of live virus and slows progress toward finding suitable treatments for infection. While vaccines from several companies have proven very effective in combating the virus, few treatments exist for those who do succumb to the viral-induced systemic disease called COVID-19. AREAS COVERED: This short review focuses on fluorescent quantum dot-based modeling of SARS-CoV-2. New BSL-2 viral models are essential for finding small molecules and biologics that may be effective in stopping viral infection, as well as treating already infected individuals. Nanoparticles are invaluable tools for biological research as they can be used to both model pathogens and serve as a platform for developing vaccines. EXPERT OPINION: Visualizing viral activity with fluorescent quantum dots enables both biochemical and cell-based assays to detect virus-host receptor interactions, cellular activity after binding to the cell plasma membrane, screening for interventions using small-molecule drug repurposing, and testing of novel biologics. Quantum dots can also be used for diagnostic assays, vaccine development, and importantly, pan-antiviral drugs to address variants that may escape the immune response.


Subject(s)
COVID-19 , Quantum Dots , Antiviral Agents/pharmacology , COVID-19/drug therapy , Drug Discovery , Humans , SARS-CoV-2
15.
Mikrochim Acta ; 188(12): 430, 2021 11 25.
Article in English | MEDLINE | ID: covidwho-1530326

ABSTRACT

Recent experience with the COVID-19 pandemic should be a lesson learnt with respect to the effort we have to invest in the development of new strategies for the treatment of viral diseases, along with their cheap, easy, sensitive, and selective detection. Since we live in a globalized world where just hours can play a crucial role in the spread of a virus, its detection must be as quick as possible. Thanks to their chemical stability, photostability, and superior biocompatibility, carbon dots are a kind of nanomaterial showing great potential in both the detection of various virus strains and a broad-spectrum antiviral therapy. The biosensing and antiviral properties of carbon dots can be tuned by the selection of synthesis precursors as well as by easy post-synthetic functionalization. In this review, we will first summarize current options of virus detection utilizing carbon dots by either electrochemical or optical biosensing approaches. Secondly, we will cover and share the up-to-date knowledge of carbon dots' antiviral properties, which showed promising activity against various types of viruses including SARS-CoV-2. The mechanisms of their antiviral actions will be further adressed as well. Finally, we will discuss the advantages and distadvantages of the use of carbon dots in the tangled battle against viral infections in order to provide valuable informations for further research and development of new virus biosensors and antiviral therapeutics.


Subject(s)
COVID-19 Testing/methods , COVID-19/diagnosis , COVID-19/drug therapy , COVID-19/therapy , Carbon/chemistry , Quantum Dots , Antiviral Agents/pharmacology , Biocompatible Materials , Biosensing Techniques , Electrochemistry , Humans , Molecular Targeted Therapy , Nanostructures , Phototherapy , Polymers , SARS-CoV-2 , Virus Diseases
16.
Biosens Bioelectron ; 198: 113810, 2022 Feb 15.
Article in English | MEDLINE | ID: covidwho-1517064

ABSTRACT

Exploring reliable and highly-sensitive SARS-CoV-2 antibody diagnosis by point-of-care (POC) manner, holds great public health significance for extensive COVID-19 screening and controlling. Unfortunately, the currently applied gold based lateral flow immunoassay (GLFIA) may expose both false-negative and false-positive interpretations owing to the sensitivity and specificity limitations, which may cause significant risk and waste of public resources for large population screening. To simultaneously overcome the drawbacks of GLFIA, a novel fluorescent LFIA based on signal amplification and dual-antigen sandwich structure was established with largely improved sensitivity and specificity. The compact three-dimensional incorporation of hydrophobic quantum dots within dendritic affinity templates and multilayer surface derivation guaranteed a high and robust fluorescence of single label, which lowered the false negative rate of GLFIA prominently. A dual-antigen sandwich structure using labeled/immobilized SARS-CoV-2 spike receptor binding domain antigen for capturing total human SARS-CoV-2 antibody was developed, instead of general indirect antibody capturing approach, to reduce the false positive rate of GLFIA. Over 300 cases of COVID-19 negative and 97 cases of COVID-19 positive samples, the current assay revealed a 100% sensitivity and 100% specificity confirmed by both polymerase chain reaction (PCR) and chemiluminescence immunoassay (CLIA), compared with the considerable misinterpretation cases by currently applied GLFIA. The quantitative results verified by receiver operating characteristic curve and other statistical analysis indicated a well-distinguished positive/negative sample groups. The proposed strategy is highly sensitive towards low concentrated SARS-CoV-2 antibody serums and highly specific towards serums from COVID-19 negative persons and patients infected by other viruses.


Subject(s)
Biosensing Techniques , COVID-19 , Quantum Dots , Antibodies, Viral , Humans , Immunoassay , SARS-CoV-2 , Sensitivity and Specificity
17.
ACS Appl Mater Interfaces ; 13(34): 40342-40353, 2021 Sep 01.
Article in English | MEDLINE | ID: covidwho-1366784

ABSTRACT

Sensitive point-of-care methods for detecting severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antigens in clinical specimens are urgently needed to achieve rapid screening of viral infection. We developed a magnetic quantum dot-based dual-mode lateral flow immunoassay (LFIA) biosensor for the high-sensitivity simultaneous detection of SARS-CoV-2 spike (S) and nucleocapsid protein (NP) antigens, which is beneficial for improving the detection accuracy and efficiency of SARS-CoV-2 infection in the point-of-care testing area. A high-performance magnetic quantum dot with a triple-QD shell (MagTQD) nanotag was first fabricated and integrated into the LFIA system to provide superior fluorescence signals, enrichment ability, and detectability for S/NP antigen testing. Two detection modes were provided by the proposed MagTQD-LFIA. The direct mode was used for rapid screening or urgent detection of suspected samples within 10 min, and the enrichment mode was used for the highly sensitive and quantitative analysis of SARS-CoV-2 antigens in biological samples without the interference of the "hook effect." The simultaneous detection of SARS-CoV-2 S/NP antigens was conducted in one LFIA strip, and the detection limits for two antigens under direct and enrichment modes were 1 and 0.5 pg/mL, respectively. The MagTQD-LFIA showed high accuracy, specificity, and stability in saliva and nasal swab samples and is an efficient tool with flexibility to meet the testing requirements for SARS-CoV-2 antigens in various situations.


Subject(s)
Antigens, Viral/analysis , Biosensing Techniques/methods , Coronavirus Nucleocapsid Proteins/analysis , SARS-CoV-2/chemistry , Spike Glycoprotein, Coronavirus/analysis , Antibodies, Immobilized/immunology , Antibodies, Monoclonal/immunology , Antigens, Viral/immunology , Coronavirus Nucleocapsid Proteins/immunology , Fluorescence , Fluorescent Dyes/chemistry , Humans , Immunoassay/methods , Limit of Detection , Magnetite Nanoparticles/chemistry , Nasopharynx/virology , Phosphoproteins/analysis , Phosphoproteins/immunology , Quantum Dots/chemistry , Saliva/virology , Sensitivity and Specificity , Spike Glycoprotein, Coronavirus/immunology
18.
Langmuir ; 37(33): 10223-10232, 2021 08 24.
Article in English | MEDLINE | ID: covidwho-1351921

ABSTRACT

Regarding the outbreak of the SARS Cov-2 virus pandemic worldwide, it seems necessary to provide new diagnostic methods to combat the virus. A fluorescence CdTe quantum dots-DNA (QDs-DNA) nanosensor was prepared for efficient detection of a specific target complementary DNA or RNA from the SARS Cov-2 virus using FRET experiment via forming a classic "sandwich" structure. The sequence of the complementary DNA (target DNA) is planned based on a substantial part of the SARS Cov-2 virus genome, and oligonucleotides of QDs-DNA nanoprobe are designed to complement it. The water-soluble CdTe QDs-DNA was prepared by replacing the thioglycolic acid (TGA) on the surface of QDs with capture DNA (thiolated DNA) through a ligand-exchange method. Subsequently, with the addition of complementary (target DNA) and quencher DNA (BHQ2-labeled DNA) into the QDs-DNA conjugates, sandwiched hybrids were formed. The resulting assembly brings the BHQ2-labeled DNA (as the acceptor), and the QDs (as the donor) into proximity, leading to quenching of fluorescence emission from the donor QDs through the FRET mechanism. In other words, a simple, highly sensitive, selective, and rapid approach was introduced to detect complementary DNA sequence from a specific part of the SARS Cov-2 virus genome with a detection limit of 2.52 × 10-9 mol L-1. Furthermore, the planned nanosensor was well used for the detection of RNA from SARS Cov-2 viruses in real samples with satisfactory analytical results, and the outcomes were compared with RT-PCR (Reverse Transcription Polymerase Chain Reaction) as the well-known standard method.


Subject(s)
Cadmium Compounds , Quantum Dots , Severe Acute Respiratory Syndrome , DNA , Humans , Spectrometry, Fluorescence , Tellurium
19.
Int J Mol Sci ; 22(8)2021 Apr 14.
Article in English | MEDLINE | ID: covidwho-1299443

ABSTRACT

Photodegradation of the aqueous solutions of acetylsalicylic acid, in the absence (ASA) and the presence of excipients (ASE), is demonstrated by the photoluminescence (PL). A shift of the PL bands from 342 and 338 nm to 358 and 361-397 nm for ASA and ASE in solid state and as aqueous solutions was reported. By exposure of the solution of ASA 0.3 M to UV light, a decrease in the PL band intensity was highlighted. This behavior was revealed for ASA in the presence of phosphate buffer (PB) having the pH equal to 6.4, 7, and 8 or by the interaction with NaOH 0.3 M. A different behavior was reported in the case of ASE. In the presence of PB, an increase in the intensity of the PL band of ASE simultaneously with a change of the ratio between the intensities of the bands at 361-364 and 394-397 nm was highlighted. The differences between PL spectra of ASA and ASE have their origin in the presence of salicylic acid (SAL). The interaction of ASE with NaOH induces a shift of the PL band at 405-407 nm. Arguments for the reaction of ASA with NaOH are shown by Raman scattering and FTIR spectroscopy.


Subject(s)
Aspirin/chemistry , Photolysis/radiation effects , Solutions/radiation effects , Water/chemistry , Aspirin/radiation effects , Cadmium Compounds/chemistry , Luminescence , Quantum Dots/chemistry , Spectrum Analysis, Raman , Ultraviolet Rays/adverse effects
20.
Nano Lett ; 21(12): 5209-5216, 2021 06 23.
Article in English | MEDLINE | ID: covidwho-1263457

ABSTRACT

The ability to rapidly diagnose, track, and disseminate information for SARS-CoV-2 is critical to minimize its spread. Here, we engineered a portable smartphone-based quantum barcode serological assay device for real-time surveillance of patients infected with SARS-CoV-2. Our device achieved a clinical sensitivity of 90% and specificity of 100% for SARS-CoV-2, as compared to 34% and 100%, respectively, for lateral flow assays in a head-to-head comparison. The lateral flow assay misdiagnosed ∼2 out of 3 SARS-CoV-2 positive patients. Our quantum dot barcode device has ∼3 times greater clinical sensitivity because it is ∼140 times more analytically sensitive than lateral flow assays. Our device can diagnose SARS-CoV-2 at different sampling dates and infectious severity. We developed a databasing app to provide instantaneous results to inform patients, physicians, and public health agencies. This assay and device enable real-time surveillance of SARS-CoV-2 seroprevalence and potential immunity.


Subject(s)
COVID-19 , Quantum Dots , Humans , Immunoassay , SARS-CoV-2 , Sensitivity and Specificity , Seroepidemiologic Studies , Smartphone
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