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1.
Chem Commun (Camb) ; 55(66): 9857-9860, 2019 Aug 25.
Artigo em Inglês | MEDLINE | ID: mdl-31364637

RESUMO

A novel fluorescent sensing platform for telomerase activity assay was developed by coupling a three-dimensional (3D) DNA walker with the MnO2 nanosheet-upconversion nanoparticle (UCNPs) complex-based fluorescence resonance energy-transfer (FRET) system.


Assuntos
Técnicas Biossensoriais/métodos , DNA/química , Transferência Ressonante de Energia de Fluorescência/instrumentação , Compostos de Manganês/química , Nanoestruturas/química , Óxidos/química , Telomerase/metabolismo , Limite de Detecção
2.
World J Microbiol Biotechnol ; 35(8): 129, 2019 Aug 02.
Artigo em Inglês | MEDLINE | ID: mdl-31376017

RESUMO

Metal whole-cell biosensors (WCBs) have been reported as very useful tools to detect and quantify the presence of bioavailable fractions of certain metals in water and soil samples. In the current work, two bacterial WCBs able to report Cr(VI) presence and plants growing on Cr(VI)-enriched soil/medium were used to assess the potential transfer of this metal to organisms of higher trophic levels, and the risk of transfer to the food chain. To do it, the functionality of the WCBs within tissues of inoculated plants in contact with Cr(VI)-contaminated soil and water was studied in vitro and in a controlled greenhouse environment. One WCB was the previously described Ochrobactrum tritici pCHRGFP2 and the second, Nitrospirillum amazonense pCHRGFP2, is a newly engineered naturally-occurring endophytic microorganism. Three rice varieties (IAC 4440, BRS 6 CHUÍ, IRGA 425) and one maize variety (1060) were tested as hosts and subjected to Cr(VI) treatments (25 µM), with different results obtained. Inoculation of each WCB into plants exposed to Cr(VI) showed GFP expression within plant tissues. WCBs penetrated the root tissues and later colonized the shoots and leaves. In general, a higher fluorescence signal was detected in roots, together with a higher Cr content and denser WCB colonization. Best fluorescence intensities per plant biomass of shoots were obtained for plant host IRGA 425. Therefore, by analyzing colonized tissues, both WCBs allowed the detection of Cr(VI) contamination in soils and its transfer to plants commonly used in crops for human diet.


Assuntos
Técnicas Biossensoriais/métodos , Cromo/análise , Ochrobactrum/crescimento & desenvolvimento , Oryza/química , Rhodospirillaceae/crescimento & desenvolvimento , Zea mays/química , Disponibilidade Biológica , Engenharia Metabólica , Microscopia de Fluorescência , Ochrobactrum/genética , Ochrobactrum/metabolismo , Oryza/crescimento & desenvolvimento , Oryza/microbiologia , Rhodospirillaceae/genética , Rhodospirillaceae/metabolismo , Poluentes do Solo/análise , Poluentes Químicos da Água/análise , Zea mays/crescimento & desenvolvimento , Zea mays/microbiologia
4.
Chem Commun (Camb) ; 55(71): 10571-10574, 2019 Aug 29.
Artigo em Inglês | MEDLINE | ID: mdl-31417999

RESUMO

A portable dual-mode sensing platform based on a self-standing TiO2 nanotube membrane is developed for simultaneously performing both qualitative analysis by the naked eye and quantitative analysis by ionic current. This dual-mode diagnosis strategy exhibits a high performance in telomerase detection in urine specimens from patients with bladder cancer.


Assuntos
Nanotubos/química , Telomerase/urina , Titânio/química , Neoplasias da Bexiga Urinária/diagnóstico , Técnicas Biossensoriais/instrumentação , Técnicas Biossensoriais/métodos , Cor , Ouro/química , Humanos , Membranas Artificiais , Nanopartículas Metálicas/química , Prata/química , Neoplasias da Bexiga Urinária/urina
5.
Chem Commun (Camb) ; 55(71): 10603-10606, 2019 Aug 29.
Artigo em Inglês | MEDLINE | ID: mdl-31424058

RESUMO

A truly ratiometric homogeneous electrochemical biosensor has been developed for sensitive miRNA detection based on the unique diffusion/intercalation properties of electroactive dyes without the need for electrode modification or materials preparation.


Assuntos
Técnicas Biossensoriais/métodos , Corantes/química , Técnicas Eletroquímicas/métodos , Substâncias Intercalantes/química , MicroRNAs/análise , DNA/química , Eletrodos , Exodesoxirribonucleases/química , Compostos Ferrosos/química , Limite de Detecção , Metalocenos/química , Azul de Metileno/química , Oxirredução
6.
Chem Commun (Camb) ; 55(71): 10615-10618, 2019 Aug 29.
Artigo em Inglês | MEDLINE | ID: mdl-31428753

RESUMO

A programmable molecular beacon (MB) with a good discrimination capability for mature and precursor microRNAs was loaded onto the surface of Mo2C quantum dots (QDs) for accurate detection of intracellular mature microRNAs.


Assuntos
Corantes Fluorescentes/química , MicroRNAs/análise , Molibdênio/química , Pontos Quânticos/química , Animais , Técnicas Biossensoriais/métodos , Linhagem Celular Tumoral , Humanos , Limite de Detecção , Camundongos , Imagem Óptica/métodos , Precursores de RNA/análise , Propriedades de Superfície
7.
Analyst ; 144(17): 5284-5291, 2019 Sep 07.
Artigo em Inglês | MEDLINE | ID: mdl-31372627

RESUMO

5,10,15,20-Tetrakis(4-carboxyl phenyl)porphyrin (Por) modified Co(OH)2 deposited on the surface of GO nanocomposites (Por/Co(OH)2/GO) were prepared and characterized by transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS), and XRD. For the first time, H2TCPP/Co(OH)2/GO is found to have enhanced peroxidase-like activity and catalyze the oxidation of the substrate 3,3,5,5-tetramethylbenzidine (TMB) by hydrogen peroxide (H2O2). Notably, the colorless TMB rapidly transformed into blue oxTMB in just 60 s, which was easily observed visually. The catalytic kinetics of H2TCPP/Co(OH)2/GO is in accord with the Michaelis-Menten equation. The catalytic mechanism of H2TCPP/Co(OH)2/GO nanocomposites is attributed to hydroxyl radicals (˙OH), due to decomposition of H2O2, which is verified by using terephthalic acid as a fluorescent probe. What's more, H2O2 can be detected in a wide linear detection range from 5 to 35 mM with a detection limit of 0.385 mM. Furthermore, based on the excellent peroxidase-like activity of H2TCPP/Co(OH)2/GO, a colorimetric sensor is established to sensitively detect glutathione (GSH) in a linear range from 10 to 300 µM with a low detection limit of 9.5 µM.


Assuntos
Cobalto/química , Grafite/química , Hidróxidos/química , Nanocompostos/química , Peroxidases/química , Porfirinas/química , Benzidinas/química , Materiais Biomiméticos , Técnicas Biossensoriais/métodos , Catálise , Glutationa/análise , Peróxido de Hidrogênio/química , Radical Hidroxila/química , Cinética , Limite de Detecção , Oxirredução , Sensibilidade e Especificidade , Propriedades de Superfície
8.
Analyst ; 144(17): 5108-5116, 2019 Sep 07.
Artigo em Inglês | MEDLINE | ID: mdl-31373337

RESUMO

We report here the influence of antibody immobilization strategy for protein immunosensors on screen printed carbon electrode arrays in terms of antibody binding activity, analytical sensitivity, limit of detection, and stability. Horseradish peroxidase (HRP) was the model analyte with anti-HRP immobilized on the sensors, and HRP activity was used for detection. Covalently immobilized anti-HRP antibodies on electrodes coated with chitosan, electrochemically reduced graphene oxide (rGO), and dense gold nanoparticle (AuNP) films had only 20-30% of the total immobilized antibodies active for binding. Active antibodies increased to 60% with passively adsorbed antibodies on bare electrodes, to 85% with oriented antibodies using protein A covalently immobilized on AuNP-coated carbon electrode, and to 98% when attached to protein A passively adsorbed onto bare electrodes. Passively adsorbed antibodies on bare electrodes lost activity in 1-2 days, but antibodies immobilized using other strategies remained relatively stable after 5 days. Covalent immobilization gave limits of detection (LOD) of 40 fg mL-1, while passively adsorbed antibodies or protein A on carbon electrodes had LODs 4-8 fg mL-1, but were unstable. Sensitivity was highest for antibodies covalently attached to AuNP electrodes (2.40 nA per log pg per mL) that also had highest antibody coverage, and decreased slightly when protein A on AuNP was used to orient antibodies. Passively adsorbed antibodies and oriented antibodies on protein A gave slightly lower sensitivities. Immobilization strategy or antibody orientation did not have a significant effect on LOD, but dynamic range increased as the number of active antibodies on sensor surfaces increased.


Assuntos
Anticorpos Imobilizados/química , Carbono/química , Técnicas Biossensoriais/métodos , Quitosana/química , Técnicas Eletroquímicas/métodos , Eletrodos , Grafite/química , Peroxidase do Rábano Silvestre/química , Imunoensaio/métodos , Limite de Detecção , Oxirredução , Propriedades de Superfície
9.
Analyst ; 144(17): 5098-5107, 2019 Sep 07.
Artigo em Inglês | MEDLINE | ID: mdl-31373344

RESUMO

Simultaneous detection and regulation of tumor-related genes presents a promising strategy for early diagnosis and treatment of cancer, but achieving this has been a huge challenge for both chemical and biomedical communities. Towards this objective, we have devised a novel aptamer-tethered, DNAzyme-embedded molecular beacon (MB) for multiple functions in cancer cells. In this design, a tumor targeting aptamer was employed to specifically deliver the sensor into cancer cells for target gene detection, and an RNA-cleaving DNAzyme was embedded to realize gene regulation. Both aptamer-tethering and DNAzyme-embedding had little influence on the sensor performance, with a detection limit of ∼2 nM and high specificity. After delivering into tumor cells, our device could monitor the tumor-related genes by producing detectable fluorescence signals, and regulate the gene expression at both mRNA and protein levels as evidenced by the RT-PCR and western blot analyses. This study provides a simple and efficient strategy to rationally combine various functional nucleic acids for multi-functional applications in living cells, which hold great potential for cancer diagnosis and therapy.


Assuntos
Aptâmeros de Nucleotídeos/química , Técnicas Biossensoriais/métodos , DNA Catalítico/química , Genes Neoplásicos , Linhagem Celular Tumoral , Humanos , Limite de Detecção
10.
J Agric Food Chem ; 67(35): 9942-9949, 2019 Sep 04.
Artigo em Inglês | MEDLINE | ID: mdl-31403785

RESUMO

We develop an ultrasensitive T2-mediated immunosensor based on the coordination chemistry and Cu(I)-catalyzed 1,3-dipolar cycloaddition of azide andalkyne (CuAAC) and apply it for the detection of pesticide residues. We functionalize polyglutamic acid (PGA) on polystyrene to form a brush-like nanostructure that has a large loading capacity of Cu(II) through the coordination chemistry between PGA and Cu(II). Such a brush-like nanostructure could be used to chelate Cu(II) to modulate the CuAAC between azide-functionalized 1000 nm polystyrene (PS1000) and alkyne-functionalized 30 nm magnetic nanoparticles (MNP30), and the MNP30-PS1000 conjugate as a product of CuAAC can act as a magnetic probe in this T2-based immunosensor. This click chemistry and coordination chemistry-mediated immunosensor allows for an ultrasensitive detection for chlorpyrifos residue (0.022 ng/mL), a 58-fold enhancement compared with that of enzyme-linked immunosorbent assay (1.28 ng/mL), providing a promising platform for detection of trace small molecules.


Assuntos
Técnicas Biossensoriais/métodos , Clorpirifos/análise , Nanoestruturas/química , Resíduos de Praguicidas/análise , Alquinos/química , Azidas/química , Técnicas Biossensoriais/instrumentação , Quelantes/química , Química Click , Cobre/química , Água Potável/análise , Lagos/análise , Ácido Poliglutâmico/química , Sensibilidade e Especificidade , Poluentes da Água/análise
11.
Chem Commun (Camb) ; 55(67): 9959-9962, 2019 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-31364996

RESUMO

On the basis of BSA stabilized tetraphenylethylene nanocrystals (BSA-TPE NCs) as aggregation-induced enhanced electrochemiluminescence (ECL) emitters with high ECL efficiency and good biocompatibility, as well as molecular recognition between ß-CD and ferrocene, an ultrasensitive and versatile ECL biosensing platform was constructed to achieve microRNA detection in cancer cells.


Assuntos
Materiais Biocompatíveis/química , Corantes Fluorescentes/química , MicroRNAs/análise , Nanopartículas/química , Soroalbumina Bovina/química , Estilbenos/química , Animais , Técnicas Biossensoriais/instrumentação , Técnicas Biossensoriais/métodos , Bovinos , Linhagem Celular Tumoral , Técnicas Eletroquímicas , Compostos Ferrosos/química , Humanos , Limite de Detecção , Medições Luminescentes , Metalocenos/química , Sensibilidade e Especificidade , beta-Ciclodextrinas/química
12.
J Agric Food Chem ; 67(32): 9104-9111, 2019 Aug 14.
Artigo em Inglês | MEDLINE | ID: mdl-31334655

RESUMO

Gold nanoflowers (GNFs) exhibit stronger light scattering ability than gold nanospheres (GNSs) with the same diameter, thereby contributing to enhancing the sensitivity of the scattering-based sensing method. However, the application of GNFs in biosensors based on dynamic light scattering (DLS) has not been yet reported. Herein, we describe for the first time an improved no-wash immunosensor based on dynamic light scattering for the detection of Escherichia coli O157:H7 (E. coli O157:H7) in milk using GNFs for sensitive signal transduction. To achieve this goal, a thiolated amphiphilic carboxyl ligand was introduced to modify the GNF surface and improve solution stability and antibody functionalization. Several key factors that affect the detection sensitivity of our developed GNF_DLS immunosensor were systematically investigated. Under the optimal conditions, our proposed GNF_DLS immunosensor provided an excellent linear detection for E. coli O157:H7 within the range from 6 × 100 to 6 × 104 colony-forming units (CFU)/mL, with a limit of detection of 2.7 CFU/mL. Combined with our previously reported two-step large-volume immunomagnetic separation (IMS) method, the designed GNF_DLS immunosensor can sensitively, selectively, and accurately detect the presence of E. coli O157:H7 in pasteurized milk. The potential of our GNF_DLS method for monitoring the presence of a single bacterial cell in 1 mL of sample solution was also demonstrated. Overall, the developed GNF_DLS immunosensor can be used for the rapid and high-sensitivity determination of pathogenic bacteria and can be extended for the ultrasensitive no-wash detection of other trace analytes.


Assuntos
Técnicas Biossensoriais/métodos , Difusão Dinâmica da Luz/métodos , Escherichia coli O157/isolamento & purificação , Leite/microbiologia , Animais , Técnicas Biossensoriais/instrumentação , Bovinos , Difusão Dinâmica da Luz/instrumentação , Escherichia coli O157/crescimento & desenvolvimento , Contaminação de Alimentos/análise , Ouro/química , Leite/química , Nanoestruturas/química , Sensibilidade e Especificidade
14.
Chem Commun (Camb) ; 55(59): 8564-8566, 2019 Jul 18.
Artigo em Inglês | MEDLINE | ID: mdl-31271158

RESUMO

Diphenylalanine (FF), as the smallest unit and core recognition motif of ß-amyloid (Aß), could self-assemble into nanofibers, which induces an early onset of Alzheimer's disease (AD). Green/near-infrared fluorescent BODIPY probes were designed and synthesized to detect FF-assembly, providing unique insights into the chemical and molecular mechanism of Aß aggregation and drug development for AD.


Assuntos
Compostos de Boro/química , Corantes Fluorescentes/química , Fenilalanina/análogos & derivados , Técnicas Biossensoriais/métodos , Compostos de Boro/síntese química , Corantes Fluorescentes/síntese química , Nanofibras/química , Fenilalanina/análise , Fenilalanina/química , Multimerização Proteica
15.
Chem Commun (Camb) ; 55(61): 8963-8966, 2019 Aug 07.
Artigo em Inglês | MEDLINE | ID: mdl-31290488

RESUMO

We develop a simple method for sensitive detection of alkaline phosphatase (ALP) based on the ligase amplification reaction-catalyzed assembly of a single quantum dot (QD)-based nanosensor. This nanosensor requires only a single ligase enzyme to achieve ultrahigh sensitivity with a detection limit of 5.63 × 10-7 U mL-1, and can be applied for kinetic analysis, inhibitor screening, and ALP measurement in cell extracts.


Assuntos
Fosfatase Alcalina/análise , Técnicas Biossensoriais/métodos , DNA Ligases/química , Ensaios Enzimáticos/métodos , Transferência Ressonante de Energia de Fluorescência/métodos , Pontos Quânticos/química , Biotina/química , Carbocianinas/química , DNA/química , DNA/genética , Sondas de DNA/química , Sondas de DNA/genética , Fluorescência , Células HEK293 , Humanos , Limite de Detecção , Células MCF-7 , Hibridização de Ácido Nucleico , Imagem Individual de Molécula , Estreptavidina/química
16.
17.
Chem Commun (Camb) ; 55(65): 9653-9656, 2019 Aug 21.
Artigo em Inglês | MEDLINE | ID: mdl-31342021

RESUMO

The fast and accurate real-time monitoring of hydrogen peroxide (H2O2) secreted from living cells plays a critical role in clinical diagnosis and management. Herein, we report low-cost and self-supported MoS2 nanosheet arrays for non-enzymatic eletrochemical H2O2 detection. Under the optimal test conditions, such MoS2 electrodes exhibit extremely promising electrocatalytic performance with a low detection limit of 1.0 µM (S/N = 3) and an excellent sensitivity of 5.3 mA mM-1 cm-2. Furthermore, the detection of the trace amount of H2O2 secreted from live A549 cancer cells was successfully performed with this biosensor.


Assuntos
Dissulfetos/química , Peróxido de Hidrogênio/análise , Molibdênio/química , Nanoestruturas/química , Células A549 , Técnicas Biossensoriais/métodos , Carbono/química , Dissulfetos/síntese química , Técnicas Eletroquímicas/métodos , Humanos , Limite de Detecção
19.
Adv Clin Chem ; 91: 31-98, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31331490

RESUMO

Specific nucleic acid detection in vitro or in vivo has become increasingly important in the discovery of genetic diseases, diagnosing pathogen infection and monitoring disease treatment. One challenge, however, is that the amount of target nucleic acid in specimens is limited. Furthermore, direct sensing methods are also unable to provide sufficient sensitivity and specificity. Fortunately, due to advances in nanotechnology and nanomaterials, nanotechnology-based bioassays have emerged as powerful and promising approaches providing ultra-high sensitivity and specificity in nucleic acid detection. This chapter presents an overview of strategies used in the development and integration of nanotechnology for nucleic acid detection, including optical and electrical detection methods, and nucleic acid assistant recycling amplification strategies. Recent 5 years representative examples are reviewed to demonstrate the proof-of-concept with promising applications for DNA/RNA detection and the underlying mechanism for detection of DNA/RNA with the higher sensitivity and selectivity. Furthermore, a brief discussion of common unresolved issues and future trends in this field is provided both from fundamental and practical point of view.


Assuntos
DNA/química , RNA/química , Técnicas Biossensoriais/métodos , Humanos , Nanotecnologia , Técnicas de Amplificação de Ácido Nucleico/métodos
20.
Environ Monit Assess ; 191(8): 507, 2019 Jul 24.
Artigo em Inglês | MEDLINE | ID: mdl-31342281

RESUMO

Glyphosate is the most widespread herbicide and its global use is steadily increasing. Although glyphosate is considered to have low toxicity, its wide application has raised concerns about its effects on human health. The extensive use of glyphosate has risen a need of its continuous monitoring in drinking and surface waters to assure in accordance with the set standards. Within the present study, we have developed a novel assay for the on-site detection of glyphosate by combining flow-through technology with the high specificity of immunorecognition. The proposed biosensing system was based on the detection of fluorescence signal generated by the quantitative replacement of glyphosate in antigen-antibody complex with IgY-type anti-glyphosate antibodies on microbeads by synthetic 5-carboxytetramethylrhodamine (5-TAMRA) conjugated glyphosate. The working range of this assay was in low millimolar range and the time required for glyphosate detection around 0.5 h. The applicability of the immunoassay for glyphosate detection in surface water was tested and the biosensor results were validated with high-performance liquid chromatography.


Assuntos
Técnicas Biossensoriais/métodos , Monitoramento Ambiental/métodos , Glicina/análogos & derivados , Herbicidas/análise , Glicina/análise , Humanos , Imunoensaio , Reprodutibilidade dos Testes , Fatores de Tempo , Poluentes Químicos da Água
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