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1.
Appl Spectrosc ; 78(9): 912-921, 2024 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-39090839

RESUMO

Growing demand for pesticides has created an environment prone to deceptive activities, where counterfeit or adulterated pesticide products infiltrate the market, often escaping rapid detection. This situation presents a significant challenge for sensor technology, crucial in identifying authentic pesticides and ensuring agricultural safety practices. Raman spectroscopy emerges as a powerful technique for detecting adulterants. Coupling the electrochemical techniques allows a more specific and selective detection and compound identification. In this study, we evaluate the efficacy of spectroelectrochemical measurements by coupling a potentiostat and Raman spectrograph to identify paraquat, a nonselective herbicide banned in several countries. Our findings demonstrate that applying -0.70 V during measurements yields highly selective Raman spectra, highlighting the primary vibrational bands of paraquat. Moreover, the selective Raman signal of paraquat was discernible in complex samples, including tap water, apple, and green cabbage, even in the presence of other pesticides such as diquat, acephate, and glyphosate. These results underscore the potential of this technique for reliable pesticide detection in diverse and complex matrices.


Assuntos
Malus , Paraquat , Análise Espectral Raman , Análise Espectral Raman/métodos , Paraquat/análise , Malus/química , Herbicidas/análise , Brassica/química , Técnicas Eletroquímicas/métodos , Contaminação de Alimentos/análise , Praguicidas/análise
2.
Chemosphere ; 361: 142481, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-38823428

RESUMO

The study presents the successful development of a new electrochemical sensor with low cost and disposability for application in nitrofurazone detection in environmental and pharmaceutical samples. The sensors were fabricated using materials obtained from local storage and conductive carbon ink. The modification of the screen-printed electrodes with the hybrid nanomaterial based on silver nanoparticles, carbon quantum dots, and carbon nanotubes showed synergistic contributions in the nitrofurazone electrooxidation, as observed in the wide linear range (0.008 at 15.051 µM), with a sensitivity of 0.650 µA/µM. The limit of detection obtained was 4.6 nM. Differential pulse voltammetry, cyclic voltammetry, X-ray photoelectron spectroscopy, X-ray diffraction analysis, and high-resolution transmission electron microscopy were used to evaluate the electrochemical and structural characteristics. Studies of possible interferences were considered with nitrofurazone in the presence of the ions and organic molecules. The results were satisfactory, with a variation of 93.3% ± 4.39% at 100% ± 2.40%. The low volume used in the analyses (50 µL), disposability, high sensibility, selectivity, and low limit of detection are advantages that make the proposed sensor an electrochemical tool of high viability for the NFZ detection in environmental matrices and pharmaceutical formulations.


Assuntos
Antibacterianos , Técnicas Eletroquímicas , Nanopartículas Metálicas , Nanotubos de Carbono , Nitrofurazona , Nitrofurazona/análise , Nitrofurazona/química , Técnicas Eletroquímicas/métodos , Nanotubos de Carbono/química , Nanopartículas Metálicas/química , Antibacterianos/análise , Limite de Detecção , Prata/química , Eletrodos , Pontos Quânticos/química
3.
Mikrochim Acta ; 190(8): 321, 2023 07 25.
Artigo em Inglês | MEDLINE | ID: mdl-37491620

RESUMO

Neglected tropical diseases are those caused by infectious agents or parasites and are considered endemic in low-income populations. These diseases also have unacceptable indicators and low investment in research, drug production, and control. Tropical diseases such as leishmaniasis are some of the main causes of morbidity and mortality around the globe. Electrochemical immunosensors are promising tools for diagnostics against these diseases. One such benefit is the possibility of assisting diagnosis in isolated regions, where laboratory infrastructure is lacking. In this work, different peptides were investigated to detect antibodies against Leishmania in human and canine serum samples. The peptides evaluated (395-KKG and 395-G) have the same recognition site but differ on their solid-binding domains, which ensure affinity to spontaneously bind to either graphene oxide (GO) or graphene quantum dots (GQD). Cyclic voltammetry and differential pulse voltammetry were employed to investigate the electrochemical behavior of each assembly step and the role of each solid-binding domain coupled to its anchoring material. The graphene affinity peptide (395-G) showed better reproducibility and selectivity when coupled to GQD. Under the optimized set of experimental conditions, negative and positive human serum samples responses were distinguished based on a cut-off value of 82.5% at a 95% confidence level. The immunosensor showed selective behavior to antibodies against Mycobacterium leprae and Mycobacterium tuberculosis, which are similar antibodies and potentially sources of false positive tests. Therefore, the use of the graphene affinity peptide as a recognition site achieved outstanding performance for the detection of Leishmania antibodies.


Assuntos
Técnicas Biossensoriais , Grafite , Leishmaniose , Animais , Cães , Humanos , Carbono/química , Grafite/química , Reprodutibilidade dos Testes , Imunoensaio , Peptídeos , Anticorpos , Leishmaniose/diagnóstico
4.
Talanta Open ; 7: 100201, 2023 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-36959870

RESUMO

To help meet the global demand for reliable and inexpensive COVID-19 testing and environmental analysis of SARS-CoV-2, the present work reports the development and application of a highly efficient disposable electrochemical immunosensor for the detection of SARS-CoV-2 in clinical and environmental matrices. The sensor developed is composed of a screen-printed electrode (SPE) array which was constructed using conductive carbon ink printed on polyethylene terephthalate (PET) substrate made from disposable soft drink bottles. The recognition site (Spike S1 Antibody (anti-SP Ab)) was covalently immobilized on the working electrode surface, which was effectively modified with carbon black (CB) and gold nanoparticles (AuNPs). The immunosensing material was subjected to a multi-technique characterization analysis using X-ray diffraction (XRD), transmission electron microscopy (TEM), and scanning electron microscopy (SEM) with elemental analysis via energy dispersive spectroscopy (EDS). The electrochemical characterization of the electrode surface and analytical measurements were performed using cyclic voltammetry (CV) and square-wave voltammetry (SWV). The immunosensor was easily applied for the conduct of rapid diagnoses or accurate quantitative environmental analyses by setting the incubation period to 10 min or 120 min. Under optimized conditions, the biosensor presented limits of detection (LODs) of 101 fg mL-1 and 46.2 fg mL-1 for 10 min and 120 min incubation periods, respectively; in addition, the sensor was successfully applied for SARS-CoV-2 detection and quantification in clinical and environmental samples. Considering the costs of all the raw materials required for manufacturing 200 units of the AuNP-CB/PET-SPE immunosensor, the production cost per unit is 0.29 USD.

5.
Mikrochim Acta ; 190(4): 136, 2023 03 15.
Artigo em Inglês | MEDLINE | ID: mdl-36920574

RESUMO

A poly(thiophene acetic acid)/Au/poly(methylene blue) nanostructured interface was electrochemically assembled step-by-step on screen-printed carbon electrodes (SPCE) for label-free detection of p53 protein. The initial electrical conductive properties of the polymeric interface were increased with an additional layer of poly(methylene blue) electropolymerized in the presence of gold nanoparticles. The nano-immunosensing architecture was prepared by covalent immobilization of anti-p53 antibodies as bioreceptors through the poly(thiophene acetic acid) moieties. The nano-immunosensor assembly was extensively characterized by ultraviolet-visible spectrophotometry, dynamic and electrophoretic light scattering, scanning electron microscopy, X-ray photoelectron spectroscopy, Raman spectroscopy, atomic force microscopy, cyclic voltammetry, and electrochemical impedance spectroscopy. Under optimal conditions, p53 was specifically and selectively detected by square wave voltammetry in a linear range between 1 and 100 ng mL-1 with a limit of detection of 0.65 ng mL-1. In addition, the electrochemical nano-immunosensor detected p53 in spiked human serum samples and colorectal cancer cell lysates, and the results were validated with a standard spectrophotometric method using a paired samples t test, which did not exhibit significant differences between both methods. The resultant p53 nano-immunosensor is simple to assemble, robust, and has the potential for point-of-care biomarker detection applications.


Assuntos
Ácido Acético , Nanopartículas Metálicas , Humanos , Técnicas Eletroquímicas/métodos , Ouro/química , Nanopartículas Metálicas/química , Azul de Metileno , Tiofenos
6.
Mikrochim Acta ; 189(6): 228, 2022 05 25.
Artigo em Inglês | MEDLINE | ID: mdl-35610491

RESUMO

A label-free nanoimmunosensor is reported based on p53/CeO2/PEDOT nanobiocomposite-decorated screen-printed gold electrodes (SPAuE) for the electrochemical detection of anti-p53 autoantibodies. CeO2 nanoparticles (NPs) were synthesized and stabilized with cyanopropyltriethoxysilane by a soft chemistry method. The nanoimmunosensing architecture was prepared by in situ electropolymerization of 3,4-ethylenedioxythiophene (EDOT) on SPAuE in the presence of CeO2 NPs. The CeO2 NPs and Ce/PEDOT/SPAuE were characterized by scanning and transmission electron microscopy, dynamic and electrophoretic light scattering, ultraviolet-visible spectrophotometry, X-ray diffraction, Fourier-transform infrared spectroscopy, cyclic voltammetry, and electrochemical impedance spectroscopy. Ce/PEDOT/SPAuE was biofunctionalized with p53 antigen by covalent bonding for the label-free determination of anti-p53 autoantibodies by differential pulse voltammetry. The nanobiocomposite-based nanoimmunosensor detected anti-p53 autoantibodies in a linear range from 10 to 1000 pg mL-1, with a limit of detection (LOD) of 3.2 pg mL-1. The nanoimmunosensor offered high specificity, selectivity, and long-term storage stability with great potential to detect anti-p53 autoantibodies in serum samples. Overall, incorporating organo-functional nanoparticles into polymeric matrices can provide a simple-to-assemble, rapid, and ultrasensitive approach for on-site screening of anti-p53 autoantibodies and other disease-related biomarkers with low sample volumes.


Assuntos
Técnicas Biossensoriais , Nanopartículas Metálicas , Nanocompostos , Autoanticorpos , Técnicas Biossensoriais/métodos , Compostos Bicíclicos Heterocíclicos com Pontes , Cério , Nanopartículas Metálicas/química , Nanocompostos/química , Polímeros
7.
Mikrochim Acta ; 189(3): 127, 2022 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-35233646

RESUMO

Smart electronic devices based on micro-controllers, also referred to as fashion electronics, have raised wearable technology. These devices may process physiological information to facilitate the wearer's immediate biofeedback in close contact with the body surface. Standard market wearable devices detect observable features as gestures or skin conductivity. In contrast, the technology based on electrochemical biosensors requires a biomarker in close contact with both a biorecognition element and an electrode surface, where electron transfer phenomena occur. The noninvasiveness is pivotal for wearable technology; thus, one of the most common target tissues for real-time monitoring is the skin. Noninvasive biosensors formats may not be available for all analytes, such as several proteins and hormones, especially when devices are installed cutaneously to measure in the sweat. Processes like cutaneous transcytosis, the paracellular cell-cell unions, or even reuptake highly regulate the solutes content of the sweat. This review discusses recent advances on wearable devices based on electrochemical biosensors for biomarkers with a complex blood-to-sweat partition like proteins and some hormones, considering the commented release regulation mechanisms to the sweat. It highlights the challenges of wearable epidermal biosensors (WEBs) design and the possible solutions. Finally, it charts the path of future developments in the WEBs arena in converging/emerging digital technologies.


Assuntos
Técnicas Biossensoriais , Dispositivos Eletrônicos Vestíveis , Biomarcadores/análise , Hormônios/análise , Suor/química
8.
Anal Chim Acta ; 1166: 338548, 2021 Jun 29.
Artigo em Inglês | MEDLINE | ID: mdl-34022998

RESUMO

Human immunodeficiency virus (HIV) is still considered a pandemic, and the detection of p24-HIV protein has an important role in the early diagnosis of HIV in adults and newborns. The accessibility of these trials depends on the price and execution difficulty of the method, which can be reduced using electrochemical methods by using enzymeless approaches, disposable and accurate devices. In this work, graphene quantum dots were acquired by a simple synthesis and employed as an electrochemical signal amplifier and support for the aptamer immobilization through a feasible and stable modification of disposable screen-printed electrodes. The device has been easily assembled and used to detect p24-HIV protein without the interference of similar proteins or sample matrix. Using the best set of experimental conditions, a linear correlation between analytical signal and log of p24-HIV concentration from 0.93 ng mL-1 to 93 µg mL-1 and a limit of detection of 51.7 pg mL-1 were observed. The developed device was applied to p24 determination in spiked human serum and provided distinct levels of signal for positive and negative samples, successfully identifying real samples with the target protein. This sensor is a step towards the development of point-of-care devices and the popularization of electrochemical methods for trials and diagnostics of relevant diseases.


Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Grafite , Infecções por HIV , Pontos Quânticos , Adulto , Técnicas Eletroquímicas , Eletrodos , Infecções por HIV/diagnóstico , Proteínas do Vírus da Imunodeficiência Humana , Humanos , Recém-Nascido , Limite de Detecção
9.
Talanta ; 224: 121804, 2021 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-33379030

RESUMO

The present work reports the development of a sensitive and selective method for ethinylestradiol detection using screen-printed electrode (SPE) modified with functionalized graphene (FG), graphene quantum dots (GQDs) and magnetic nanoparticles coated with molecularly imprinted polymers (mag@MIP). The performance of the mag@MIP sensor was compared with that of a non-molecularly imprinted sensor (mag@NIP). Chemical and physical characterizations of the mag@NIP and mag@MIP sensors were performed using scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDS) and Brunauer-Emmett-Teller (BET) techniques. The electrochemical behavior of the electrodes investigated, which included (mag@MIP)-GQDs-FG-NF/SPE, (mag@NIP)-GQDs-FG-NF/SPE, GQDs-FG-NF/SPE and FG-NF/SPE, was evaluated by cyclic voltammetry. The results obtained show a significant increase in peak current magnitude for (mag@MIP)-GQDs-FG-NF/SPE. Using square wave voltammetry experiments, the efficiency of the (mag@MIP)-GQDs-FG-NF/SPE sensor was also tested under optimized conditions. The linear response range obtained for ethinylestradiol concentration was 10 nmol L-1 to 2.5 µmol L-1, with limit of detection of 2.6 nmol L-1. The analytical signal of the (mag@MIP)-GQDs-FG-NF/SPE sensor suffered no interference from different compounds and the sensor exhibited good repeatability. The proposed sensor was successfully applied for ethynilestradiol detection in river water, serum and urine samples, where recovery rates between 96 to 105% and 97-104% were obtained for environmental and biological samples, respectively.


Assuntos
Grafite , Nanopartículas de Magnetita , Impressão Molecular , Pontos Quânticos , Técnicas Eletroquímicas , Eletrodos , Etinilestradiol , Limite de Detecção , Polímeros Molecularmente Impressos
10.
J Forensic Sci ; 65(6): 2121-2128, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32602943

RESUMO

The need for agile and proper identification of drugs of abuse has encouraged the scientific community to improve and to develop new methodologies. The drug lysergic acid diethylamide (LSD) is still widely used due to its hallucinogenic effects. The use of voltammetric methods to analyze narcotics has increased in recent years, and the possibility of miniaturizing the electrochemical equipment allows these methods to be applied outside the laboratory; for example, in crime scenes. In addition to portability, the search for affordable and sustainable materials for use in electroanalytical research has grown in recent decades. In this context, employing paper substrate, graphite pencil, and silver paint to construct paper-based electrodes is a great alternative. Here, a paper-based device comprising three electrodes was drawn on 300 g/m2 watercolor paper with 8B pencils, and its efficiency was compared to the efficiency of a commercially available screen-printed carbon electrode. Square wave voltammetry was used for LSD analysis in aqueous medium containing 0.05 mol/L LiClO4 . The limits of detection and quantification were 0.38 and 1.27 µmol/L, respectively. Both electrodes exhibited a similar voltammetric response, which was also confirmed during analysis of a seized LSD sample, with recovery of less than 10%. The seized samples were previously analyzed by GCMS technique, employing the full scan spectra against the software spectral library. The electrode selectivity was also tested against 3,4-methylenedioxymethamphetamine (MDMA) and methamphetamine. It was possible to differentiate these compounds from LSD, indicating that the developed paper-based device has potential application in forensic chemistry analyses.


Assuntos
Eletroquímica/instrumentação , Eletrodos , Alucinógenos/análise , Dietilamida do Ácido Lisérgico/análise , Papel , Toxicologia Forense/instrumentação , Humanos , Limite de Detecção , Metanfetamina/isolamento & purificação , N-Metil-3,4-Metilenodioxianfetamina/isolamento & purificação
11.
Biosens Bioelectron ; 163: 112238, 2020 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-32568700

RESUMO

This work reports a new sensitive strategy for the determination of tau protein, a hallmark of Alzheimer's disease (AD), involving a sandwich immunoassay and amperometric detection at disposable screen-printed carbon electrodes (SPCEs) modified with a gold nanoparticles-poly(amidoamine) (PAMAM) dendrimer nanocomposite (3D-Au-PAMAM) covalently immobilized onto electrografted p-aminobenzoic acid (p-ABA). The capture antibody (CAb) was immobilized by crosslinking with glutaraldehyde (GA) on the amino groups of the 3D-Au-PAMAM-p-ABA-SPCE, where tau protein was sandwiched with a secondary antibody labeled with horseradish peroxidase (HRP-DAb). Amperometry at -200 mV (vs the Ag pseudo-reference electrode) upon the addition of hydroquinone (HQ) as electron transfer mediator and H2O2 as the enzyme substrate was used to detect the immunocomplex formation. The great analytical performance of the immunosensor in terms of selectivity and low limit of detection (LOD) (1.7 pg mL-1) allowed the direct determination of the target protein in raw plasma samples and in brain tissue extracts from healthy individuals and post mortem diagnosed AD patients, using a simple and fast protocol.


Assuntos
Doença de Alzheimer , Técnicas Biossensoriais , Nanopartículas Metálicas , Doença de Alzheimer/diagnóstico , Encéfalo , Carbono , Técnicas Eletroquímicas , Eletrodos , Ouro , Humanos , Peróxido de Hidrogênio , Imunoensaio , Limite de Detecção , Proteínas tau
12.
J Environ Manage ; 266: 110587, 2020 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-32392140

RESUMO

Urine and struvite are promising organic fertilizers that can replace conventional fertilizers. However, these fertilizers can have some emerging contaminants, such as dipyrone. This drug is one of the main painkillers consumed in the world and its continuous and indiscriminate intake can promote the camouflage of symptoms of other diseases, anaphylactic shock and even death. Thus, a fast, sensitive, inexpensive and portable method for metamizole (dipyrone) determination in several matrices, applied as organic fertilizers, has been successfully developed using portable equipment and bare carbon screen-printed electrodes in conjunction with square wave voltammetry (SWV). The main SWV operating parameters were optimized (equilibrium time (60 s), step potential (6 mV), modulation amplitude (50 mV) and frequency (10 Hz)) using univariate experiments. The proposed method presented a limit of detection of 0.097 ± 0.002 µmol L-1 (RSD = 2.72%, n = 3) for dipyrone in 0.1 mol L-1 HCl and R2 equal to 0.993. The determination in the struvite sample presented a concentration of 0.47 µmol L-1 of dipyrone. Urine sample used in the production of struvite and urine collected from an individual 10h after ingestion of 500 mg dipyrone tablet showed concentrations of 15.2 and 590 µmol L-1 of dipyrone, respectively. The recovery test in fortified struvite sample showed values between 91 and 102% (RSD = 3.1%, n = 3) and of 102% (SD = 3.7%, n = 3) in human urine, indicating that there is no matrix effect. These results reinforce the possibility of applying the proposed method on-site in a practical and fast way, without the need of significant amounts of sample promoting a more sustainable chemistry.


Assuntos
Dipirona , Fertilizantes , Carbono , Eletrodos , Humanos , Estruvita
13.
Talanta ; 207: 120277, 2020 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-31594622

RESUMO

A low-cost and disposable microcell was constructed with a screen-printed electrode for the non-enzymatic electrochemical determination of creatinine. The working electrode was modified with carbon black and maintained in contact with paper-adsorbed iron (III) ions. A small sample volume of 3 µL was required for the device operation. Then, iron (III) ions were complexed in the presence of creatinine in a chemical step, followed by an electrochemical reduction of non-complexed metallic ions in excess. Cyclic voltammetry and differential-pulse voltammetry experiments were employed for the electrochemical characterizations and analytical performance evaluation of the microcell. The working electrode modification with carbon black provided a significant increase of analytical signal. The sensor presented a linear response for creatinine concentrations ranging from 0.10 to 6.5 mmol L-1, with a limit of detection of 0.043 mmol L-1. Experiments for creatinine determination in real samples were successful performed through of standard recovery in urine.


Assuntos
Creatinina/análise , Eletroquímica/instrumentação , Análise em Microsséries/instrumentação , Impressão , Creatinina/química , Creatinina/urina , Eletrodos , Química Verde , Humanos , Ferro/química , Limite de Detecção , Fuligem/química
14.
Anal Bioanal Chem ; 411(25): 6667-6676, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31384983

RESUMO

Human epidermal growth factor receptor-2 (HER2) is an important biomarker in the diagnosis and prognosis of breast cancer. This work aimed to develop an aptasensor to detect HER2 in human serum. HER2 aptamer was immobilized by electrostatic adsorption on the surface of a homemade screen-printed electrode modified with poly-L-lysine. Measurements were made by differential pulse voltammetry using methylene blue as a redox indicator. A calibration curve was constructed (R2 = 0.997) using different concentrations of HER2 protein (10-60 ng/mL) in PBS buffer (pH 7.4), with a detection limit of 3.0 ng/mL. The aptasensor showed good reproducibility with relative standard deviations (RSDs) of 3% and remained stable for 3 days with an RSD around 2%. When the tests were performed with serum from a healthy woman, a peak of 6.72 µA was found without the addition of the protein. When we tested the serum of a woman with HER2+ breast cancer, we obtained a signal of 2.65 µA; the same pattern was found when adding to protein in serum control, i.e., the higher the concentration of protein, the lower the signal. The aptasensor was characterized by scanning electron microscopy and isothermal titration calorimetry (ITC), showing excellent interaction between aptamer and target protein. The results revealed a promising and sensitive tool capable of detecting HER2 protein in human serum with albumin depletion, aiding in the molecular diagnosis of breast cancer. Graphical abstract.


Assuntos
Aptâmeros de Nucleotídeos/química , Neoplasias da Mama/sangue , Receptor ErbB-2/sangue , Técnicas Biossensoriais/instrumentação , Técnicas Biossensoriais/métodos , Neoplasias da Mama/diagnóstico , Detecção Precoce de Câncer , Técnicas Eletroquímicas/instrumentação , Técnicas Eletroquímicas/métodos , Eletrodos , Desenho de Equipamento , Feminino , Humanos , Limite de Detecção , Receptor ErbB-2/análise , Reprodutibilidade dos Testes
15.
Talanta ; 205: 120110, 2019 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-31450419

RESUMO

Tristeza is a disease that affects citrus crops in general, caused by the Citrus tristeza virus (CTV). It is considered an economically important virus diseases in citrus, which is present in the main citrus producing regions all around the world. Early detection of CTV is crucial to avoid any epidemics and substantial economic losses for the citrus growers. Consequently, the development of rapid, accurate, and sensitive methods capable of detecting the virus in the early stages of the disease is highly desired. Based on that, a low-cost and rapid magneto-immunoassay methodology to detect the capsid protein from CTV (CP-CTV) was proposed. For this, magnetic beads were decorated with antibodies anti-CP-CTV and horseradish peroxidase enzyme (HRP) and applied for the capture and separation of CP-CTV from the sample solutions. The magnetically captured biomarker was detected using a simple disposable microfluidic electrochemical device (DµFED) constructed by rapid prototyping technique and composed by an array of immunosensors. In DµFED, the electrodes were modified with monoclonal antibody anti-CP-CTV and the detection was carried out using amperometry, based on the hydroquinone/H2O2 catalytic redox reaction due to the presence of HRP label in an immune-sandwich structure. The proposed immunoassay presented excellent linearity with a wide linear range of concentration of 1.95-10.0 × 103 fg mL-1 and ultralow detection limit of 0.3 fg mL-1. The disposable device was successfully applied for the detection of Citrus tristeza virus in healthy and infected plant samples, where it showed good agreements with the comparative method of enzyme-linked immunosorbent assay (ELISA). The developed immunoassay methodology showed a sensitive and selective way in the detection of CTV. Hence, it can be considered as a promising analytical alternative for rapid and low-cost diagnosis of Tristeza disease in citrus.


Assuntos
Closterovirus/isolamento & purificação , Dispositivos Lab-On-A-Chip , Animais , Anticorpos Imobilizados/imunologia , Anticorpos Monoclonais Murinos/imunologia , Proteínas do Capsídeo/análise , Proteínas do Capsídeo/imunologia , Citrus/virologia , Técnicas Eletroquímicas/instrumentação , Técnicas Eletroquímicas/métodos , Eletrodos , Ouro/química , Imunoensaio/métodos , Separação Imunomagnética/métodos , Limite de Detecção , Nanopartículas Metálicas/química , Camundongos , Técnicas Analíticas Microfluídicas/instrumentação , Técnicas Analíticas Microfluídicas/métodos , Doenças das Plantas/virologia , Folhas de Planta/virologia , Reprodutibilidade dos Testes
16.
Anal Chim Acta ; 1033: 49-57, 2018 Nov 29.
Artigo em Inglês | MEDLINE | ID: mdl-30172331

RESUMO

This work presents potential applications of low-cost fused deposition modeling 3D-printers to fabricate multiuse 3D-printed electrochemical cells for flow or batch measurements as well as the 3D-printing of electrochemical sensing platforms. Electrochemical cells and sensors were printed with acrylonitrile butadiene styrene (ABS) and conductive graphene-doped polylactic acid (G-PLA) filaments, respectively. The overall printing operation time and estimated cost per cell were 6 h and $ 6.00, respectively, while the sensors were printed within minutes (16 sensor strips of 1 × 2 cm in 10 min at a cost of $ 1.00 each sensor). The cell performance is demonstrated for the amperometric detection of tert-butylhydroquinone, dipyrone, dopamine and diclofenac by flow-injection analysis (FIA) and batch-injection analysis (BIA) using different working electrodes, including the proposed 3D-printed sensor, which presented comparable electroanalytical performance with other carbon-based electrodes (LOD of 0.1 µmol L-1 for dopamine). Raman spectroscopy and scanning electron microscopy of the 3D-printed sensor indicated the presence of graphene nanoribbons within the polymeric matrix. Electrochemical impedance spectroscopy and heterogeneous electron transfer constants (k0) for the redox probe Ru(NH3)6+3 revealed that a glassy-carbon electrode presented faster electron transfer rates than the 3D-printed sensor; however, the latter presented lower LOD values for dopamine and catechol probably due to oxygenated functional groups at the G-PLA surface.


Assuntos
Técnicas Eletroquímicas , Análise de Injeção de Fluxo , Impressão Tridimensional , Diclofenaco/análise , Dipirona/análise , Dopamina/análise , Transporte de Elétrons , Hidroquinonas/análise , Microscopia Eletrônica de Varredura , Análise Espectral Raman , Propriedades de Superfície
17.
Biosens Bioelectron ; 100: 577-582, 2018 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-29031228

RESUMO

Juvenile idiopathic arthritis (JIA) is a wide group of diseases, characterized by synovial inflammation and joint tissue damage. Due to the delay in the implementation of biomarkers into clinical practice and the association with severe sequels, there is an imperative need for new JIA diagnosis strategies. Electrochemical biosensors based on screen-printed electrodes and peptides are promising alternatives for molecular diagnosis. In this work, a novel biosensor for detecting juvenile idiopathic arthritis (JIA) was developed based on the immobilization of the PRF+1 mimetic peptide, as recognition biological element, on the surface of screen-printed carbon electrode. This biosensor was able to discriminate the JIA positive and negative serum samples from different individuals using differential pulse voltammetry, presenting limits of detection and quantification in diluted samples of 1:784 (v/v) and 1:235 (v/v), respectively. Evaluation by electrochemical impedance spectroscopy showed RCT 3 times higher for JIA positive sample than for a pool of human serum samples from healthy individuals. Surface analysis of the biosensor by atomic force microscopy, after contact with JIA positive serum, presented great globular clusters irregularly distributed. The long-term stability of the biosensor was evaluated, remaining functional for over 40 days of storage (after storage at 8°C). Therefore, a simple, miniaturized and selective biosensor was developed, being the first one based on mimetic peptide and screen-printed carbon electrode, aiming at the diagnosis of the juvenile idiopathic arthritis in real serum samples.


Assuntos
Artrite Juvenil/diagnóstico , Técnicas Biossensoriais/métodos , Peptídeos/química , Artrite Juvenil/sangue , Técnicas Biossensoriais/instrumentação , Espectroscopia Dielétrica , Técnicas Eletroquímicas/instrumentação , Técnicas Eletroquímicas/métodos , Eletrodos , Desenho de Equipamento , Humanos , Modelos Moleculares
18.
Biosens Bioelectron ; 99: 156-162, 2018 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-28755608

RESUMO

A novel fully disposable microfluidic electrochemical array device (µFED) was developed and successfully applied for detection of the biomarker estrogen receptor alpha (ERα). The µFED was constructed using low-cost materials and an inexpensive home cutter printer enabled the manufacture of dozens of µFEDs in less than 2h, at a cost of less than US$ 0.20 in material per device. The µFED incorporates counter and reference electrodes and eight carbon-based working electrodes, which were modified with DNA sequences known as estrogen response elements (DNA-ERE), where ERα binds specifically. Paramagnetic particles heavily decorated with anti-ERα antibody and horseradish peroxidase (MP-Ab-HRP) were used to efficiently capture ERα from the sample solution. The ERα-MP-Ab-HRP bioconjugate formed was injected into the µFED and incubated with the DNA-ERE-modified electrodes, followed by amperometric detection with application of -0.2V vs. Ag|AgCl while a mixture of H2O2 and hydroquinone was injected into the microfluidic device. An ultralow limit of detection of 10.0 fg mL-1 was obtained with the proposed method. The performance of the assay, in terms of sensitivity and reproducibility, was studied using undiluted calf serum, and excellent recoveries in the range of 94.7-108% were achieved for the detection of ERα in MCF-7 cell lysate. The µFED system can be easily constructed and applied for multiplex biomarker detection, making the device an excellent cost-effective alternative for cancer diagnosis, especially in developing countries.


Assuntos
Biomarcadores Tumorais/genética , Técnicas Biossensoriais , Neoplasias da Mama/diagnóstico , Receptor alfa de Estrogênio/isolamento & purificação , Neoplasias da Mama/genética , Receptor alfa de Estrogênio/genética , Humanos , Limite de Detecção , Células MCF-7
19.
ACS Appl Mater Interfaces ; 9(33): 27433-27440, 2017 Aug 23.
Artigo em Inglês | MEDLINE | ID: mdl-28742317

RESUMO

Breast cancer is the most common cancer in women worldwide. The detection of biomarkers has played a significant role in the early diagnosis and prognosis of breast cancer. Herein, we describe the construction of a disposable microfluidic immunoarray device (DµID) for the rapid and low-cost detection of CA15-3 (carbohydrate antigen 15-3), a protein biomarker for breast cancer. The DµID was constructed using a simple and rapid prototyping technique and was applied to detect CA15-3 in cancer patients. The DµID construction was based on the use of a double-sided adhesive card with a microfluidic channel and a screen-printed array with 8 electrodes. Both the immunoarray and microfluidic channel were designed using an inexpensive home cutter printer and using low-cost materials. The immunoarray was modified using the layer-by-layer technique aiming at immobilizing the primary antibody. For the biomarker detection, magnetic particles (MPs) modified with polyclonal antibodies and peroxidase enzymes were used as a strategy for capture, separation, and preconcentration of the biomarker, in addition to amplification of the electroanalytical signal. The preconcentration and amplification strategies integrated with the nanostructured immunosensors of the DµID meaningfully contributed toward the detection of CA15-3 with a limit of detection (LoD) of 6 µU mL-1, requiring as low as 2 µL of serum samples for 8 simultaneous detections. The obtained LoD was 1200 times lower compared to those of other immunosensors previously reported in the literature. The DµID was applied for the detection of CA15-3 in real samples of breast cancer patients and was found to present an excellent correlation with the well-established commercial electrochemiluminescence immunoassay. The association of the DµID with nanostructured surfaces and analyte capturing with bioconjugated paramagnetic particles is essentially a promising breakthrough for the low-cost and accurate detection of cancer biomarkers.


Assuntos
Microfluídica , Biomarcadores Tumorais , Técnicas Biossensoriais , Humanos , Imunoensaio , Dispositivos Lab-On-A-Chip , Limite de Detecção
20.
Anal Chim Acta ; 934: 1-8, 2016 08 31.
Artigo em Inglês | MEDLINE | ID: mdl-27506338

RESUMO

This work presents the potential application of organic-resistant screen-printed graphitic electrodes (SPGEs) for fuel analysis. The required analysis of the antioxidant 2,6-di-tert-butylphenol (2,6-DTBP) in biodiesel and jet fuel is demonstrated as a proof-of-concept. The screen-printing of graphite, Ag/AgCl and insulator inks on a polyester substrate (250 µm thickness) resulted in SPGEs highly compatible with liquid fuels. SPGEs were placed on a batch-injection analysis (BIA) cell, which was filled with a hydroethanolic solution containing 99% v/v ethanol and 0.1 mol L(-1) HClO4 (electrolyte). An electronic micropipette was connected to the cell to perform injections (100 µL) of sample or standard solutions. Over 200 injections can be injected continuously without replacing electrolyte and SPGE strip. Amperometric detection (+1.1 V vs. Ag/AgCl) of 2,6-DTBP provided fast (around 8 s) and precise (RSD = 0.7%, n = 12) determinations using an external calibration curve. The method was applied for the analysis of biodiesel and aviation jet fuel samples and comparable results with liquid and gas chromatographic analyses, typically required for biodiesel and jet fuel samples, were obtained. Hence, these SPGE strips are completely compatible with organic samples and their combination with the BIA cell shows great promise for routine and portable analysis of fuels and other organic liquid samples without requiring sophisticated sample treatments.

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