Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 2.925
Filtrar
1.
Food Chem ; 367: 130727, 2022 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-34371276

RESUMO

An electrochemical sensor was fabricated by modifying nanoporous gold (NPG)-coated glassy carbon electrode (NPG/GCE) with functionalized graphene oxide /chitosan/ionic liquid nanocomposites (fGO/CS/IL). The introduction of ionic liquid (IL) and chitosan (CS) induced higher dispersibility of functionalized graphene oxide (fGO), and was beneficial for the combination of fGO/CS/IL with NPG/GCE. As a result of the synergistic effect of NPG and fGO/CS/IL, the resulted functionalized graphene oxide/chitosan/ionic liquid nanocomposites/nanoporous gold /glassy carbon electrode (fGO/CS/IL/NPG/GCE) showed the highest redox peak current response signal of Amaranth (E123) due to ultrahigh surface area, electronic conductivity as well as the improvement of the surface structure. Under optimized conditions, the enhanced peak currents represented excellent analytical performance for detection of Amaranth in the concentration range from 8.0 to 1200.0 nM. Meanwhile, the fGO/CS/IL/NPG/GCE presented satisfactory sensitivity and selectivity, excellent reproducibility, and long-time stability. For practical applications, the fGO/CS/IL/NPG/GCE was validated for the determination of Amaranth in three types of drinks with satisfactory results.


Assuntos
Técnicas Biossensoriais , Quitosana , Grafite , Líquidos Iônicos , Nanocompostos , Nanoporos , Técnicas Eletroquímicas , Eletrodos , Ouro , Reprodutibilidade dos Testes
2.
BMC Genomics ; 22(1): 726, 2021 Oct 07.
Artigo em Inglês | MEDLINE | ID: mdl-34620074

RESUMO

BACKGROUND: The golden lion tamarin (Leontopithecus rosalia) is an endangered Platyrrhine primate endemic to the Atlantic coastal forests of Brazil. Despite ongoing conservation efforts, genetic data on this species remains scarce. Complicating factors include limitations on sample collection and a lack of high-quality reference sequences. Here, we used nanopore adaptive sampling to resequence the L. rosalia mitogenome from feces, a sample which can be collected non-invasively. RESULTS: Adaptive sampling doubled the fraction of both host-derived and mitochondrial sequences compared to sequencing without enrichment. 258x coverage of the L. rosalia mitogenome was achieved in a single flow cell by targeting the unfinished genome of the distantly related emperor tamarin (Saguinus imperator) and the mitogenome of the closely related black lion tamarin (Leontopithecus chrysopygus). The L. rosalia mitogenome has a length of 16,597 bp, sharing 99.68% sequence identity with the L. chrysopygus mitogenome. A total of 38 SNPs between them were identified, with the majority being found in the non-coding D-loop region. DNA methylation and hydroxymethylation were directly detected using a neural network model applied to the raw signal from the MinION sequencer. In contrast to prior reports, DNA methylation was negligible in mitochondria in both CpG and non-CpG contexts. Surprisingly, a quarter of the 642 CpG sites exhibited DNA hydroxymethylation greater than 1% and 44 sites were above 5%, with concentration in the 3' side of several coding regions. CONCLUSIONS: Overall, we report a robust new mitogenome assembly for L. rosalia and direct detection of cytosine base modifications in all contexts.


Assuntos
Genoma Mitocondrial , Leontopithecus , Nanoporos , Animais , DNA , Epigenoma , Fezes
3.
Nat Commun ; 12(1): 5811, 2021 10 04.
Artigo em Inglês | MEDLINE | ID: mdl-34608151

RESUMO

Chemical reactions of single molecules, caused by rapid formation or breaking of chemical bonds, are difficult to observe even with state-of-the-art instruments. A biological nanopore can be engineered into a single molecule reactor, capable of detecting the binding of a monatomic ion or the transient appearance of chemical intermediates. Pore engineering of this type is however technically challenging, which has significantly restricted further development of this technique. We propose a versatile strategy, "programmable nano-reactors for stochastic sensing" (PNRSS), by which a variety of single molecule reactions of hydrogen peroxide, metal ions, ethylene glycol, glycerol, lactic acid, vitamins, catecholamines or nucleoside analogues can be observed directly. PNRSS presents a refined sensing resolution which can be further enhanced by an artificial intelligence algorithm. Remdesivir, a nucleoside analogue and an investigational anti-viral drug used to treat COVID-19, can be distinguished from its active triphosphate form by PNRSS, suggesting applications in pharmacokinetics or drug screening.


Assuntos
Técnicas Biossensoriais/instrumentação , Nanoporos , Inteligência Artificial , Processos Estocásticos
4.
Nano Lett ; 21(19): 8236-8243, 2021 10 13.
Artigo em Inglês | MEDLINE | ID: mdl-34597051

RESUMO

Graphene oxide (GO) is receiving tremendous attention in membrane separation; however, its desalination performances remain suboptimal because of excessive swelling and tortuous transport pathways. Herein, we chemically joint GO nanosheets and phenolic nanomeshes together to form laminated membranes comprising through-plane nanopores and stabilized nanochannels. GO and phenolic/polyether nanosheets are mixed to form stacked structures and then treated in H2SO4 to remove polyether to produce nanomeshes and to chemically joint GO with phenolic nanomeshes. Thus-synthesized laminated membranes possess enhanced interlayer interactions and narrowed interlayer spacings down to 6.4 Å. They exhibit water permeance up to 165.6 L/(m2 h bar) and Na2SO4 rejection of 97%, outperforming most GO-based membranes reported so far. Moreover, the membranes are exceptionally stable in water because the chemically jointed laminates suppress the swelling of GO. This work reports hybrid laminated structures of GO and phenolic nanomeshes, which are highly desired in desalination and other applications.


Assuntos
Grafite , Nanoporos , Membranas Artificiais , Água
5.
BMC Genomics ; 22(1): 732, 2021 Oct 09.
Artigo em Inglês | MEDLINE | ID: mdl-34627155

RESUMO

BACKGROUND: Enzyme-based host depletion significantly improves the sensitivity of clinical metagenomics. Recent studies found that real-time adaptive sequencing of DNA molecules was achieved using a nanopore sequencing machine, which enabled effective enrichment of microbial sequences. However, few studies have compared the enzyme-based host depletion and nanopore adaptive sequencing for microbial enrichment efficiency. RESULTS: To compare the host depletion and microbial enrichment efficiency of enzyme-based and adaptive sequencing methods, the present study collected clinical samples from eight children with respiratory tract infections. The same respiratory samples were subjected to standard methods, adaptive sequencing methods, enzyme-based host depletion methods, and the combination of adaptive sequencing and enzyme-based host depletion methods. We compared the host depletion efficiency, microbial enrichment efficiency, and pathogenic microorganisms detected between the four methods. We found that adaptive sequencing, enzyme-based host depletion and the combined methods significantly enriched the microbial sequences and significantly increased the diversity of microorganisms (p value < 0.001 for each method compared to standard). The highest microbial enrichment efficiency was achieved using the combined method. Compared to the standard method, the combined method increased the microbial reads by a median of 113.41-fold (interquartile range 23.32-327.72, maximum 1812), and the number of genera by a median of 70-fold (interquartile range 56.75-86.75, maximum 164). The combined method detected 6 pathogens in 4 samples with a median read of 547, compared to 5 pathogens in 4 samples with a median read of 4 using the standard method. CONCLUSION: The combined method is an effective, easy-to-run method for enriching microbial sequences in clinical metagenomics from sputum and bronchoalveolar lavage fluid samples and may improve the sensitivity of clinical metagenomics for other host-derived clinical samples.


Assuntos
Sequenciamento por Nanoporos , Nanoporos , Criança , DNA , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Metagenômica
6.
Phys Rev Lett ; 127(13): 138103, 2021 Sep 24.
Artigo em Inglês | MEDLINE | ID: mdl-34623840

RESUMO

Nanopores in 2D materials are highly desirable for DNA sequencing, yet achieving single-stranded DNA (ssDNA) transport through them is challenging. Using density functional theory calculations and molecular dynamics simulations we show that ssDNA transport through a pore in monolayer hexagonal boron nitride (h-BN) is marked by a basic nanomechanical conflict. It arises from the notably inhomogeneous flexural rigidity of ssDNA and causes high friction via transient DNA desorption costs exacerbated by solvation effects. For a similarly sized pore in bilayer h-BN, its self-passivated atomically smooth edge enables continuous ssDNA transport. Our findings shed light on the fundamental physics of biopolymer transport through pores in 2D materials.


Assuntos
Compostos de Boro/química , DNA de Cadeia Simples/química , Nanoestruturas/química , Fenômenos Biofísicos , Modelos Químicos , Simulação de Dinâmica Molecular , Nanoporos
7.
J Phys Chem Lett ; 12(37): 9132-9141, 2021 Sep 23.
Artigo em Inglês | MEDLINE | ID: mdl-34523927

RESUMO

Different nucleotides generate specific ionic currents that discriminate between the nucleotides while they are passing through the nanopore constriction. MspA is a commonly used nanopore for DNA sequencing. However, the reasons of the current variation remain ambiguous. Our work unveils the microscopic mechanism of current variation for an ssDNA passing through the MspA nanopore by all-atom molecular dynamic simulations. Besides the physical rigidity and dimensions of the nucleotides, nucleotide orientation is observed to induce nonignorable current variation. Besides the generally considered MspA nanopore constriction, it is also found that the region below constriction could be used to detect and differentiate single nucleotides when the single-stranded DNA translocates in the form of base-constriction-base meshing and ratcheting across the nanopore constriction compared to other regions. The work provides a novel insight into facilitating the development of low-cost and high-throughput nanopore DNA sequencing.


Assuntos
Nanoporos , Porinas/química , Análise de Sequência de DNA/métodos , DNA de Cadeia Simples/química , DNA de Cadeia Simples/metabolismo , Simulação de Dinâmica Molecular , Mycobacterium smegmatis/metabolismo , Porinas/metabolismo
8.
Planta ; 254(4): 83, 2021 Sep 24.
Artigo em Inglês | MEDLINE | ID: mdl-34559312

RESUMO

MAIN CONCLUSION: Engineered nanocarriers have great potential to deliver different genetic cargos to plant cells and increase the efficiency of plant genetic engineering. Genetic engineering has improved the quality and quantity of crops by introducing desired DNA sequences into the plant genome. Traditional transformation strategies face constraints such as low transformation efficiency, damage to plant tissues, and genotype dependency. Smart nanovehicle-based delivery is a newly emerged method for direct DNA delivery to plant genomes. The basis of this new approach of plant genetic transformation, nanomaterial-mediated gene delivery, is the appropriate protection of transferred DNA from the nucleases present in the cell cytoplasm through the nanocarriers. The conjugation of desired nucleic acids with engineered nanocarriers can solve the problem of genetic manipulation in some valuable recalcitrant plant genotypes. Combining nano-enabled genetic transformation with the new and powerful technique of targeted genome editing, CRISPR (clustered regularly interspaced short palindromic repeats), can create new protocols for efficient improvement of desired plants. Silica-based nanoporous materials, especially mesoporous silica nanoparticles (MSNs), are currently regarded as exciting nanoscale platforms for genetic engineering as they possess several useful properties including ordered and porous structure, biocompatibility, biodegradability, and surface chemistry. These specific features have made MSNs promising candidates for the design of smart, controlled, and targeted delivery systems in agricultural sciences. In the present review, we discuss the usability, challenges, and opportunities for possible application of nano-enabled biomolecule transformation as part of innovative approaches for target delivery of genes of interest into plants.


Assuntos
Sistemas CRISPR-Cas , Nanoporos , Sistemas CRISPR-Cas/genética , Produtos Agrícolas/genética , Edição de Genes , Engenharia Genética , Genoma de Planta
9.
Nanoscale ; 13(32): 13658-13664, 2021 Aug 28.
Artigo em Inglês | MEDLINE | ID: mdl-34477641

RESUMO

Multiplexing methods which are capable of measurement of multiple analytes in a single assay are of great importance in many fields. The conventional strategy for simultaneous detection of multiple species is to construct a sensor array. Herein, we report an innovative multiplex multi-analyte detection platform in a non-array format for protease measurement. By monitoring protease degradation of a single peptide substrate containing two cleavage sites for a disintegrin and metalloproteinase 10 (ADAM10) and a disintegrin and metalloproteinase 10 (ADAM17) in a single nanopore, simultaneous detection and quantification of these two model proteases in mixture samples could satisfactorily be accomplished. Our developed multiplexing sensing platform has the potential to be coupled with the traditional sensor array to further improve the multiplexing capability of the sensor, which may find useful applications in clinical diagnosis and prognosis.


Assuntos
Técnicas Biossensoriais , Nanoporos , Endopeptidases , Peptídeo Hidrolases , Peptídeos
10.
Nat Commun ; 12(1): 5348, 2021 09 09.
Artigo em Inglês | MEDLINE | ID: mdl-34504071

RESUMO

Single-molecule counting is the most accurate and precise method for determining the concentration of a biomarker in solution and is leading to the emergence of digital diagnostic platforms enabling precision medicine. In principle, solid-state nanopores-fully electronic sensors with single-molecule sensitivity-are well suited to the task. Here we present a digital immunoassay scheme capable of reliably quantifying the concentration of a target protein in complex biofluids that overcomes specificity, sensitivity, and consistency challenges associated with the use of solid-state nanopores for protein sensing. This is achieved by employing easily-identifiable DNA nanostructures as proxies for the presence ("1") or absence ("0") of the target protein captured via a magnetic bead-based sandwich immunoassay. As a proof-of-concept, we demonstrate quantification of the concentration of thyroid-stimulating hormone from human serum samples down to the high femtomolar range. Further optimization to the method will push sensitivity and dynamic range, allowing for development of precision diagnostic tools compatible with point-of-care format.


Assuntos
Biomarcadores/sangue , Imunoensaio/métodos , Nanoporos , Nanotecnologia/métodos , Tireotropina/sangue , Algoritmos , Proteínas Sanguíneas/análise , DNA/química , Humanos , Medicina de Precisão/métodos , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
11.
Front Cell Infect Microbiol ; 11: 696669, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34485177

RESUMO

Background: Emerging long reads sequencing technology has greatly changed the landscape of whole-genome sequencing, enabling scientists to contribute to decoding the genetic information of non-model species. The sequences generated by PacBio or Oxford Nanopore Technology (ONT) be assembled de novo before further analyses. Some genome de novo assemblers have been developed to assemble long reads generated by ONT. The performance of these assemblers has not been completely investigated. However, genome assembly is still a challenging task. Methods and Results: We systematically evaluated the performance of nine de novo assemblers for ONT on different coverage depth datasets. Several metrics were measured to determine the performance of these tools, including N50 length, sequence coverage, runtime, easy operation, accuracy of genome and genomic completeness in varying depths of coverage. Based on the results of our assessments, the performances of these tools are summarized as follows: 1) Coverage depth has a significant effect on genome quality; 2) The level of contiguity of the assembled genome varies dramatically among different de novo tools; 3) The correctness of an assembled genome is closely related to the completeness of the genome. More than 30× nanopore data can be assembled into a relatively complete genome, the quality of which is highly dependent on the polishing using next generation sequencing data. Conclusion: Considering the results of our investigation, the advantage and disadvantage of each tool are summarized and guidelines of selecting assembly tools are provided under specific conditions.


Assuntos
Nanoporos , Genoma , Genômica , Sequenciamento de Nucleotídeos em Larga Escala , Análise de Sequência de DNA , Tecnologia
12.
Nano Lett ; 21(19): 8393-8400, 2021 10 13.
Artigo em Inglês | MEDLINE | ID: mdl-34542296

RESUMO

The outbreak of the SARS-CoV-2 caused the disease COVID-19 to spread globally. Specific and sensitive detection of SARS-CoV-2 facilitates early intervention and prevents the disease from spreading. Here, we present a solid-state CRISPR-Cas12a-assisted nanopore (SCAN) sensing strategy for the specific detection of SARS-CoV-2. We introduced a nanopore-sized counting method to measure the cleavage ratio of reporters, which is used as a criterion for positive/negative classification. A kinetic cleavage model was developed and validated to predict the reporter size distributions. The model revealed the trade-offs between sensitivity, turnaround time, and false-positive rate of the SARS-CoV-2 SCAN. With preamplification and a 30 min CRISPR Cas12a assay, we achieved excellent specificity against other common human coronaviruses and a limit of detection of 13.5 copies/µL (22.5 aM) of viral RNA at a confidence level of 95%. These results suggested that the SCAN could provide a rapid, sensitive, and specific analysis of SARS-CoV-2.


Assuntos
COVID-19 , Nanoporos , Sistemas CRISPR-Cas/genética , Humanos , Técnicas de Amplificação de Ácido Nucleico , SARS-CoV-2 , Sensibilidade e Especificidade
13.
J Hazard Mater ; 416: 125976, 2021 08 15.
Artigo em Inglês | MEDLINE | ID: mdl-34492884

RESUMO

The molecular-scale adsorption mechanism of heavy metal ions in the interlayer and nanopore regions of montmorillonite (MMT) were investigated by molecular dynamics simulations. Three typical heavy metals (zinc, cadmium, and lead) were selected as the model ions, and two types of MMT (Arizona and Wyoming) were considered. The results showed that Cd2+ and Pb2+ can form both inner- and outer-sphere complexes on Wyoming MMT, while Zn2+ only formed outer-sphere complex due to the stronger hydration interaction of Zn2+ than Cd2+ and Pb2+. For Arizona MMT, all of the three ions only formed outer-sphere complexes on its interlayer and external basal surface in which the cations remained a fully hydrated state. The calculated diffusion coefficients of three cations in interlayer and nanopore indicated that their diffusion abilities were significantly impaired, implying that MMT adsorbents have a strong ability to fix and retard heavy metal ions. The derived results and mechanisms are instrumental to a profound understanding of the transport and retention of heavy metal elements in subsurface environments, and provide guidance for the management of heavy metal pollution.


Assuntos
Metais Pesados , Nanoporos , Adsorção , Bentonita , Cádmio , Íons , Simulação de Dinâmica Molecular
14.
ACS Nano ; 15(9): 14419-14429, 2021 09 28.
Artigo em Inglês | MEDLINE | ID: mdl-34583465

RESUMO

Temporal changes in electrical resistance of a nanopore sensor caused by translocating target analytes are recorded as a sequence of pulses on current traces. Prevalent algorithms for feature extraction in pulse-like signals lack objectivity because empirical amplitude thresholds are user-defined to single out the pulses from the noisy background. Here, we use deep learning for feature extraction based on a bi-path network (B-Net). After training, the B-Net acquires the prototypical pulses and the ability of both pulse recognition and feature extraction without a priori assigned parameters. The B-Net is evaluated on simulated data sets and further applied to experimental data of DNA and protein translocation. The B-Net results are characterized by small relative errors and stable trends. The B-Net is further shown capable of processing data with a signal-to-noise ratio equal to 1, an impossibility for threshold-based algorithms. The B-Net presents a generic architecture applicable to pulse-like signals beyond nanopore currents.


Assuntos
Aprendizado Profundo , Nanoporos
15.
Anal Chem ; 93(38): 13054-13062, 2021 09 28.
Artigo em Inglês | MEDLINE | ID: mdl-34519478

RESUMO

Probe-modified nanopores/nanochannels are one of the most advanced sensors because the probes interact strongly with ions and targets in nanoconfinement and create a sensitive and selective ionic signal. Recently, ionic signals have been demonstrated to be sensitive to the probe-target interaction on the outer surface of nanopores/nanochannels, which can offer more open space for target recognition and signal conversion than nanoconfined cavities. To enhance the ionic signal, we investigated the effect of grafting density, a critical parameter of the sensing interface, of the probe on the outer surface of nanochannels on the change rate of the ionic signal before and after target recognition (ß). Electroneutral peptide nucleic acids and negatively charged DNA are selected as probes and targets, respectively. The experimental results showed that when adding the same number of targets, the ß value increased with the probe grafting density on the outer surface. A theoretical model with clearly defined physical properties of each probe and target has been established. Numerical simulations suggest that the decrease of the background current and the aggregation of targets at the mouth of nanochannels with increasing probe grafting density contribute to this enhancement. This work reveals the signal mechanism of probe-target recognition on the outer surface of nanochannels and suggests a general approach to the nanochannel/nanopore design leading to sensitivity improvement on the basis of relatively good selectivity.


Assuntos
Nanoporos , Ácidos Nucleicos Peptídicos , DNA , Íons , Modelos Teóricos
16.
Nanoscale ; 13(36): 15352-15361, 2021 Sep 23.
Artigo em Inglês | MEDLINE | ID: mdl-34498657

RESUMO

Protein sequencing is essential to unveil the mechanism of cellular processes that govern the function of living organisms, and which play a crucial role in the field of drug design and molecular diagnostics. Nanopores have been proved to be effective tools in single molecule sensing, but the fast translocation speed of a peptide through a nanopore is one of the major obstacles that hinders the development of nanopore-based protein sequencing. In this work, by using molecular dynamics simulations (MDS) it is found that the peptide containing more hydrophobic residues permeates slower through a molybdenum disulfide nanopore, which originates from the strong interaction between the membrane surface and the hydrophobic residues. The binding affinity is remarkable especially for benzenoid residues as they contain a hydrophobic aromatic ring that is composed of relatively non-polar C-C and C-H bonds. By tuning the fraction of benzenoid residues of the peptide, the velocity of the protein translocation through the nanopore is well controlled. The peptide with all the hydrophobic residues being benzenoid residues is found to translocate through the nanopore almost ten times slower than the one without any benzenoid residues, which is beneficial for gathering adequate information for precise amino acid identification.


Assuntos
Nanoporos , Sequência de Aminoácidos , Simulação de Dinâmica Molecular , Transporte Proteico , Proteínas
17.
Elife ; 102021 09 28.
Artigo em Inglês | MEDLINE | ID: mdl-34581669

RESUMO

High-throughput genomics of SARS-CoV-2 is essential to characterize virus evolution and to identify adaptations that affect pathogenicity or transmission. While single-nucleotide variations (SNVs) are commonly considered as driving virus adaption, RNA recombination events that delete or insert nucleic acid sequences are also critical. Whole genome targeting sequencing of SARS-CoV-2 is typically achieved using pairs of primers to generate cDNA amplicons suitable for next-generation sequencing (NGS). However, paired-primer approaches impose constraints on where primers can be designed, how many amplicons are synthesized and requires multiple PCR reactions with non-overlapping primer pools. This imparts sensitivity to underlying SNVs and fails to resolve RNA recombination junctions that are not flanked by primer pairs. To address these limitations, we have designed an approach called 'Tiled-ClickSeq', which uses hundreds of tiled-primers spaced evenly along the virus genome in a single reverse-transcription reaction. The other end of the cDNA amplicon is generated by azido-nucleotides that stochastically terminate cDNA synthesis, removing the need for a paired-primer. A sequencing adaptor containing a Unique Molecular Identifier (UMI) is appended to the cDNA fragment using click-chemistry and a PCR reaction generates a final NGS library. Tiled-ClickSeq provides complete genome coverage, including the 5'UTR, at high depth and specificity to the virus on both Illumina and Nanopore NGS platforms. Here, we analyze multiple SARS-CoV-2 isolates and clinical samples to simultaneously characterize minority variants, sub-genomic mRNAs (sgmRNAs), structural variants (SVs) and D-RNAs. Tiled-ClickSeq therefore provides a convenient and robust platform for SARS-CoV-2 genomics that captures the full range of RNA species in a single, simple assay.


Assuntos
Sequência de Bases , Coronavirus/genética , Genoma Viral , RNA , SARS-CoV-2/genética , COVID-19/virologia , DNA Complementar , Biblioteca Gênica , Genômica , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Nanoporos , Reação em Cadeia da Polimerase , RNA Mensageiro , RNA Viral/genética , Recombinação Genética , Sequenciamento Completo do Genoma
18.
Biosens Bioelectron ; 194: 113588, 2021 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-34474277

RESUMO

Ultrasensitive, versatile sensors for molecular biomarkers are a critical component of disease diagnostics and personalized medicine as the COVID-19 pandemic has revealed in dramatic fashion. Integrated electrical nanopore sensors can fill this need via label-free, direct detection of individual biomolecules, but a fully functional device for clinical sample analysis has yet to be developed. Here, we report amplification-free detection of SARS-CoV-2 RNAs with single molecule sensitivity from clinical nasopharyngeal swab samples on an electro-optofluidic chip. The device relies on optically assisted delivery of target carrying microbeads to the nanopore for single RNA detection after release. A sensing rate enhancement of over 2,000x with favorable scaling towards lower concentrations is demonstrated. The combination of target specificity, chip-scale integration and rapid detection ensures the practicality of this approach for COVID-19 diagnosis over the entire clinically relevant concentration range from 104-109 copies/mL.


Assuntos
Técnicas Biossensoriais , COVID-19 , Nanoporos , Teste para COVID-19 , Humanos , Pinças Ópticas , Pandemias , RNA Viral/genética , SARS-CoV-2
19.
Biosens Bioelectron ; 194: 113602, 2021 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-34481241

RESUMO

Inhibition of HIV-1 protease (PR) activity is realized by exposure to 60Co γ-radiation. The radiation effects on enzyme kinetics of HIV-1 PR are subsequently monitored using nanopore sensor. Substantial loss of proteolytic efficiency towards GagPol polypeptide is observed due to the radiation treatment. Results shows ~50% of GagPol polypeptide was not involved in HIV-1 PR proteolysis by exposure to ultra-low intensity of γ-radiation (0.1K Gy), and the values reach to over 90% with high γ-ray treatment. Besides, the inactivation effect is also verified in blood samples which contain the virus protease. Our finding provides the potential benefits of γ-radiation to inactivate viral proteinic function, and might be a complementary to the designation of HIV-1 PR inhibitors.


Assuntos
Técnicas Biossensoriais , HIV-1 , Nanoporos , Protease de HIV , Proteólise
20.
ACS Sens ; 6(8): 3133-3143, 2021 08 27.
Artigo em Inglês | MEDLINE | ID: mdl-34406743

RESUMO

We report a simple method for tailoring the size of in-plane nanopores fabricated in thermoplastics for single-molecule sensing. The in-plane pores were fabricated via nanoimprint lithography (NIL) from resin stamps, which were generated from Si masters. We could reduce the size of the in-plane nanopores from 30 to ∼10 nm during the thermal fusion bonding (TFB) step, which places a cover plate over the imprinted polymer substrate under a controlled pressure and temperature to form the relevant nanofluidic devices. Increased pressures during TFB caused the cross-sectional area of the in-plane pore to be reduced. The in-plane nanopores prepared with different TFB pressures were utilized to detect single-λ-DNA molecules via resistive pulse sensing, which showed a higher current amplitude in devices bonded at higher pressures. Using this method, we also show the ability to tune the pore size to detect single-stranded (ss) RNA molecules and single ribonucleotide adenosine monophosphate (rAMP). However, due to the small size of the pores required for detection of the ssRNA and rAMPs, the surface charge arising from carboxylate groups generated during O2 plasma oxidation of the surfaces of the nanopores to make them wettable had to be reduced to allow translocation of coions. This was accomplished using EDC/NHS coupling chemistry and ethanolamine. This simple modification chemistry increased the event frequency from ∼1 s-1 to >136 s-1 for an ssRNA concentration of 100 nM.


Assuntos
Nanoporos , DNA , Nanotecnologia , Polímeros , Temperatura
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...