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1.
Talanta ; 274: 125998, 2024 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-38574541

RESUMO

Artificial nanozymes (enzyme-mimics), specifically metallic nanomaterials, have garnered significant attention recently due to their reduced preparation cost and enhanced stability in a wide range of environments. The present investigation highlights, for the first time, a straightforward green synthesis of biogenic platinum nanoparticles (PtNPs) from a natural resource, namely Prunella vulgaris (Pr). To demonstrate the effectiveness of the phytochemical extract as an effective reducing agent, the PtNPs were characterized by various techniques such as UV-vis spectroscopy, High-resolution Transmission electron microscopy (HR-TEM), zeta-potential analysis, Fourier-transform infrared spectroscopy (FTIR), and Energy dispersive spectroscopy (EDS). The formation of PtNPs with narrow size distribution was verified. Surface decoration of PtNPs was demonstrated with multitudinous functional groups springing from the herbal extract. To demonstrate their use as viable nanozymes, the peroxidase-like activity of Pr/PtNPs was evaluated through a colorimetric assay. Highly sensitive visual detection of H2O2 with discrete linear ranges and a low detection limit of 3.43 µM was demonstrated. Additionally, peroxidase-like catalytic activity was leveraged to develop a colorimetric platform to quantify glutamate biomarker levels with a high degree of selectivity, the limit of detection (LOD) being 7.00 µM. The 2,2-Diphenyl-1-picrylhydrazyl (DPPH) test was used to explore the scavenging nature of the PtNPs via the degradation of DPPH. Overall, the colorimetric assay developed using the Pr/PtNP nanozymes in this work could be used in a broad spectrum of applications, ranging from biomedicine and food science to environmental monitoring.


Assuntos
Antioxidantes , Ácido Glutâmico , Peróxido de Hidrogênio , Nanopartículas Metálicas , Platina , Prunella , Platina/química , Nanopartículas Metálicas/química , Peróxido de Hidrogênio/química , Peróxido de Hidrogênio/análise , Antioxidantes/química , Antioxidantes/análise , Antioxidantes/farmacologia , Prunella/química , Ácido Glutâmico/análise , Ácido Glutâmico/química , Extratos Vegetais/química
2.
Food Chem ; 450: 139320, 2024 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-38640530

RESUMO

4(5)-methylimidazole (4-MeI) is a potential carcinogen widely used in food colours. EU regulations specify a maximum allowable concentration of 200 ppm for 4-MeI in caramel colours. This study reports an electrochemical determination technique for 4-MeI in caramel colours for the first time. The effect of pH and interference from air were studied to optimize the detection conditions on a glassy carbon electrode in aqueous alkaline solutions using square wave voltammetry (SWV) technique. The concentration of 4-MeI was quantitatively measured down to 10 µM (∼0.8 ppm). Traditional methods such as HPLC, GC, spectrometry and immunoassays involve either expensive instrumentation and reagents or time consuming preparation and detection processes. This study demonstrates the possibility of rapid and simple electrochemical determination of (4-MeI) in food colours with minimum workup using a portable potentiostat.


Assuntos
Técnicas Eletroquímicas , Imidazóis , Imidazóis/química , Imidazóis/análise , Técnicas Eletroquímicas/instrumentação , Corantes de Alimentos/análise , Corantes de Alimentos/química , Contaminação de Alimentos/análise , Concentração de Íons de Hidrogênio , Carboidratos
3.
Biosens Bioelectron ; 246: 115857, 2024 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-38029708

RESUMO

This study unveils the electrochemically-enhanced nanozymatic activity exhibited by borophene during the reaction of 3,3',5,5'-tetramethylbenzidine (TMB) and H2O2. Herein, the surface of the pristine borophene was first modified with the addition of thiocyanate groups to improve hydroxyl radical (•OH) scavenging activity. Then, the oxidation reaction of TMB was accelerated under applied electrochemical potential. Both factors significantly improved the detection limit and drastically decreased the detection time. DPPH testing revealed that the radical scavenging nature of borophene was more than 70%, boosting its catalytic activity. In the presence of H2O2, borophene catalyzed the oxidation of TMB and produced a blue-colored solution that was linearly correlated with the concentration of H2O2 and allowed for the detection of H2O2 up to 38 nM. The present finding was further extended to nanozymatic detection of tetracyclines (TCs) using a target-specific aptamer, and the results were colorimetrically quantifiable up to 1 µM with a LOD value of 150 nM. Moreover, transferring the principles of the discussed detection method to form a portable and disposable paper-based system enabled the quantification of TCs up to 0.2 µM. All the sensing experiments in this study indicate that the nanozymatic activity of borophene has significantly improved under electrochemical potential compared to conventional nanozyme-based colorimetric detection. Hence, the present discovery of electrochemically-enhanced nanozymatic activity would be promising for various sensitive and time-dependent colorimetric sensor development initiatives in the future.


Assuntos
Técnicas Biossensoriais , Peróxido de Hidrogênio , Técnicas Biossensoriais/métodos , Antibacterianos , Tetraciclina , Tetraciclinas , Colorimetria/métodos , Peroxidase
4.
ACS Appl Mater Interfaces ; 15(24): 29425-29439, 2023 Jun 21.
Artigo em Inglês | MEDLINE | ID: mdl-37279206

RESUMO

Hospital-acquired (nosocomial) infections account for the majority of adverse health effects during care delivery, placing an immense financial strain on healthcare systems around the world. For the first time, the present article provides evidence of a straightforward pollution-free technique to fabricate a heteroatom-doped carbon dot immobilized fluorescent biopolymer composite for the development of functional textiles with antioxidant and antimicrobial properties. A simple, facile, and eco-friendly approach was devised to prepare heteroatom-doped carbon dots from waste green tea and a biopolymer. The carbon dots showed an excitation-dependent emission behavior, and the XPS data unveiled that they are co-doped with nitrogen and sulfur. A facile physical compounding strategy was adopted to fabricate a carbon dot reinforced biopolymeric composite followed by immobilization onto the textile. The composite textiles revealed excellent antioxidant activity, determined by 1,1-diphenyl-2-picrylhydrazyl (>80%) and 2,2'-azinobis-3-ethylbenzothiazoline-6-sulfonic acid assays (>90%). The results of the disc diffusion assay indicated that the composite textiles substantially inhibited the growth of both tested bacteria Escherichia coli and Bacillus subtilis with increasing coating cycles. The time-dependent antibacterial experiments revealed that the nanocomposite can inhibit significant bacterial growth within a few hours. The present study could open up the possibility for the commercialization of inexpensive smart textile substrates for the prevention of microbial contamination used for the medical and healthcare field.


Assuntos
Anti-Infecciosos , Antioxidantes , Antibacterianos/farmacologia , Antibacterianos/química , Anti-Infecciosos/farmacologia , Antioxidantes/farmacologia , Carbono/química , Corantes , Têxteis/microbiologia
5.
J Biol Eng ; 17(1): 33, 2023 Apr 27.
Artigo em Inglês | MEDLINE | ID: mdl-37106392

RESUMO

BACKGROUND: Norovirus is a common pathogen that causes foodborne outbreaks every year and the increasing number of deaths caused by it has become a substantial concern in both developed and underdeveloped countries. To date, no vaccines or drugs are able to control the outbreak, highlighting the importance of finding specific, and sensitive detection tools for the viral pathogen. Current diagnostic tests are limited to public health laboratories and/or clinical laboratories and are time-consuming. Hence, a rapid and on-site monitoring strategy for this disease is urgently needed to control, prevent and raise awareness among the general public. RESULTS: The present study focuses on a nanohybridization technique to build a higher sensitivity and faster detection response to norovirus-like particles (NLPs). Firstly, the wet chemical-based green synthesis of fluorescent carbon quantum dots and gold nanoparticles (Au NPs) has been reported. Then, a series of characterization studies were conducted on the synthesized carbon dots and Au NPs, for example, high-resolution transmission emission microscopy, fluorescence spectroscopy, fluorescence life-lime measurement, UV-visible spectroscopy, and X-ray diffraction (XRD). The fluorescence emission of the as-synthesized carbon dots and the absorption of Au NPs were located at 440 nm and 590 nm, respectively. Then, the plasmonic properties of Au NPs were utilized to enhance the fluorescence emission of carbon dots in the presence of NLPs in human serum. Here, the enhanced fluorescence response was linearly correlated up to 1 µg mL-1. A limit of detection (LOD) value was calculated to be 80.3 pg mL-1 demonstrating that the sensitivity of the proposed study is 10 times greater than that of the commercial diagnostic kits. CONCLUSIONS: The proposed exciton-plasmon interaction-based NLPs-sensing strategy was highly sensitive, specific, and suitable for controlling upcoming outbreaks. Most importantly, the overall finding in the article will take the technology a step further to applicable point-of-care (POC) devices.

6.
Foods ; 11(9)2022 Apr 19.
Artigo em Inglês | MEDLINE | ID: mdl-35563912

RESUMO

The concentration of thiocyanate (SCN-) in bodily fluids is a good indicator of potential and severe health issues such as nasal bleeding, goiters, vertigo, unconsciousness, several inflammatory diseases, and cystic fibrosis. Herein, a visual SCN- sensing method has been developed using the enzyme-like nature of positively charged gold quantum dots (Au QDs) mixed with 3,3',5,5'-tetramethylbenzidine (TMB) and hydrogen peroxide (H2O2). This research also reports a new method of synthesizing positively charged Au QDs directly from gold nanoparticles through a hydrothermal process. Microscopic imaging has showed that the Au QDs were 3-5 nm in size, and the emission wavelength was at 438 nm. Au QDs did not display any enzyme-like nature while mixed up with TMB and H2O2. However, the nanozymatic activity of Au QDs appeared when SCN- was included, leading to a very low detection limit (LOD) of 8 nM and 99-105% recovery in complex media. The steady-state kinetic reaction of Au QDs showed that Au QDs had a lower Michaelis-Menten constant (Km) toward H2O2 and TMB, which indicates that the Au QDs had a higher affinity for H2O2 and TMB than horseradish peroxidase (HRP). A mechanism study has revealed that the scavenging ability of hydroxyl (•OH) radicals by the SCN- group plays an important role in enhancing the sensitivity in this study. The proposed nanozymatic "Off-On" SCN- sensor was also successfully validated in commercial milk samples.

7.
Talanta ; 236: 122863, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34635245

RESUMO

Cannabis is currently one of the most consumed drugs in many countries. Δ9-tetrahydrocannabinol (THC) is the principal psychoactive component of this drug and is present in saliva after consumption. This paper reports a novel biomolecule-free electrochemical approach to detect an ultra-low level of THC in saliva using modified electrodes with molecules of the same analyte (THC) that are detected later via square wave voltammetry. The results from this research revealed that the electrodeposition of THC on the working electrode (sensor analyte) could highly enhance the limit of detection by improving the affinity of the THC molecules present in the sample (sample analyte) to the sensing electrode surface. Detailed descriptions about the optimization of the sensor and its performance in simple media, such as PBS, and complex media, such as simulated and real saliva, are provided. This novel and yet simple electrochemical-based sensing strategy allowed for a low limit of detection of 1.6 ng/mL THC in simulated and real saliva, distinguishing concentrations ranging from 2 to 25 ng/mL, making this technology viable for a real-world application such as roadside testing.


Assuntos
Cannabis , Alucinógenos , Dronabinol , Eletrodos , Saliva
8.
Food Chem ; 346: 128893, 2021 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-33387835

RESUMO

The excessive use of tetracyclines (TCs), a bacteriostaticantibiotic, in food products, has led to the accumulation of TCs residues in the human body, affecting human health seriously. Therefore, the development of a highly sensitive method to detect TCs in food is of utmost importance. This study reports a novel sensing strategy using aptamer-induced fluorescence fluctuation of graphene quantum dots (GQDs) and palladium nanoparticles (Pd NPs) for the rapid and label-free detection of tetracycline with a limit of detection of 45 ng.mL-1. A novel single-step synthesis of positively charged Pd NPs and one-step green synthesis of GQDs directly from graphite has been developed. The proposed strategy provides an efficient way to detect low traces of TCs and a new technique for the development of aptamer-based sensors.


Assuntos
Aptâmeros de Nucleotídeos/química , Técnicas Biossensoriais/métodos , Grafite/química , Leite/química , Paládio/química , Pontos Quânticos/química , Tetraciclina/análise , Animais , Corantes Fluorescentes/química , Humanos , Limite de Detecção , Nanopartículas Metálicas/química , Tetraciclina/química
9.
Heliyon ; 4(8): e00766, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30186985

RESUMO

A synthetic way of chiral zirconium quantum dots (Zr QDs) was presented for the first time using L(+)-ascorbic acid acts as a surface as well as chiral ligands. Different spectroscopic and microscopic analysis was performed for thorough characterization of Zr QDs. As-synthesized QDs exhibited fluorescence and circular dichroism properties, and the peaks were located at 412 nm and 352 nm, respectively. MTT assay was performed to test the cytotoxicity of the synthesized Zr QDs against rat brain glioma C6 cells. Synthesized QDs was further conjugated with anti-infectious bronchitis virus (IBV) antibodies of coronavirus to form an immunolink at the presence of the target analyte and anti-IBV antibody-conjugated magneto-plasmonic nanoparticles (MPNPs). The fluorescence properties of immuno-conjugated QD-MP NPs nanohybrids through separation by an external magnetic field enabled biosensing of coronavirus with a limit of detection of 79.15 EID/50 µL.

10.
ACS Appl Mater Interfaces ; 10(15): 12534-12543, 2018 Apr 18.
Artigo em Inglês | MEDLINE | ID: mdl-29595253

RESUMO

Rapid and sensitive detection of influenza virus is of soaring importance to prevent further spread of infections and adequate clinical treatment. Herein, an ultrasensitive colorimetric assay called magnetic nano(e)zyme-linked immunosorbent assay (MagLISA) is suggested, in which silica-shelled magnetic nanobeads (MagNBs) and gold nanoparticles are combined to monitor influenza A virus up to femtogram per milliliter concentration. Two essential strategies for ultrasensitive sensing are designed, i.e., facile target separation by MagNBs and signal amplification by the enzymelike activity of gold nanozymes (AuNZs). The enzymelike activity was experimentally and computationally evaluated, where the catalyticity of AuNZ was tremendously stronger than that of normal biological enzymes. In the spiked test, a straightforward linearity was presented in the range of 5.0 × 10-15-5.0 × 10-6g·mL-1 in detecting the influenza virus A (New Caledonia/20/1999) (H1N1). The detection limit is up to 5.0 × 10-12 g·mL-1 only by human eyes, as well as up to 44.2 × 10-15 g·mL-1 by a microplate reader, which is the lowest record to monitor influenza virus using enzyme-linked immunosorbent assay-based technology as far as we know. Clinically isolated human serum samples were successfully observed at the detection limit of 2.6 PFU·mL-1. This novel MagLISA demonstrates, therefore, a robust sensing platform possessing the advances of fathomable sample separation, enrichment, ultrasensitive readout, and anti-interference ability may reduce the spread of influenza virus and provide immediate clinical treatment.


Assuntos
Imunoadsorventes/química , Ensaio de Imunoadsorção Enzimática , Ouro , Humanos , Vírus da Influenza A Subtipo H1N1 , Vírus da Influenza A , Nanopartículas Metálicas
11.
Glob Chall ; 2(4): 1700071, 2018 Apr 26.
Artigo em Inglês | MEDLINE | ID: mdl-31565328

RESUMO

Molybdenum disulfide (MoS2), a type of transition metal dichalcogenide material, has emerged as an important class among 2D systems. When 2D MoS2 materials are reduced to 0D quantum dots (QDs), they introduce new optical properties that point to several potential technological advantages in electronic, magnetic, optical, and catalytic properties. In this study, a simple way to produce chiral MoS2 QDs from MoS2 nanopowder is presented using l(+)-ascorbic acid as a reducing agent. The calculated quantum yield of QDs is 11.06%. Experimental results reveal that the size of QDs is uniformly monodispersed (2-3 nm) and have a blue emissive fluorescence peak and circular dichroism (CD) peak located at 420 and 330 nm, respectively. Furthermore, a dual-mode detection system based on fluorescence and chirality is performed using as-synthesized MoS2 QDs, where QDs are conjugated with anti-hemagglutinin antibodies of avian influenza virus and made into an immunobridge in the presence of target virus and anti-neuraminidase antibodies conjugated magnetic nanoparticles (MNPs). The photoluminescence and CD spectra of unconjugated QDs after separated magnetochirofluorescent (MNPs-QDs) nanohybrids by external magnets enables influenza virus A (H5N1) detection with the limit of detection value of 7.35 and 80.92 pg mL-1, respectively.

12.
Biosens Bioelectron ; 103: 45-53, 2018 Apr 30.
Artigo em Inglês | MEDLINE | ID: mdl-29278812

RESUMO

An optoelectronic sensor is a rapid diagnostic tool that allows for an accurate, reliable, field-portable, low-cost device for practical applications. In this study, template-free In situ gold nanobundles (Au NBs) were fabricated on an electrode for optoelectronic sensing of fowl adenoviruses (FAdVs). Au NB film was fabricated on carbon electrodes working area using L(+) ascorbic acid, gold chroloauric acid and poly-l-lysine (PLL) through modified layer-by-layer (LbL) method. A scanning electron microscopic (SEM) image of the Au NBs revealed a NB-shaped Au structure with many kinks on its surface, which allow local electric field enhancement through light-matter interaction with graphene quantum dots (GQDs). Here, GQDs were synthesized through an autoclave-assisted method. Characterization experiments revealed blue-emissive, well-dispersed GQDs that were 2-3nm in size with the fluorescence emission peak of GQDs located at 405nm. Both Au NBs and GQDs were conjugated with target FAdVs specific antibodies that bring them close to each other with the addition of target FAdVs through antibody-antigen interaction. At close proximity, light-matter interaction between Au NBs and QDs produces a local electric signal enhancement under Ultraviolet-visible (UV-visible) light irradiation that allows the detection of very low concentrations of target virus even in complex biological media. A proposed optoelectronic sensor showed a linear relationship between the target FAdVs and the electric signal up to 10 Plaque forming unit (PFU)/mL with a limit of detection (LOD) of 8.75 PFU/mL. The proposed sensing strategy was 100 times more sensitive than conventional ELISA method.


Assuntos
Adenoviridae/isolamento & purificação , Técnicas Biossensoriais , Nanopartículas Metálicas/química , Ouro , Grafite/química , Pontos Quânticos/química
13.
Nanotheranostics ; 1(3): 272-295, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29071193

RESUMO

Outbreaks of foodborne diseases related to fresh produce have been increasing in North America and Europe. Viral foodborne pathogens are poorly understood, suffering from insufficient awareness and surveillance due to the limits on knowledge, availability, and costs of related technologies and devices. Current foodborne viruses are emphasized and newly emerging foodborne viruses are beginning to attract interest. To face current challenges regarding foodborne pathogens, a point-of-care (POC) concept has been introduced to food testing technology and device. POC device development involves technologies such as microfluidics, nanomaterials, biosensors and other advanced techniques. These advanced technologies, together with the challenges in developing foodborne virus detection assays and devices, are described and analysed in this critical review. Advanced technologies provide a path forward for foodborne virus detection, but more research and development will be needed to provide the level of manufacturing capacity required.

14.
Nanotheranostics ; 1(3): 338-345, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29071197

RESUMO

Nanomaterial-based artificial enzymes or nanozymes exhibit superior properties such as stability, cost effectiveness and ease of preparation in comparison to conventional enzymes. However, the lower catalytic activity of nanozymes limits their sensitivity and thereby practical applications in the bioanalytical field. To overcome this drawback, herein we propose a very simple but highly sensitive, specific and low-cost dual enhanced colorimetric immunoassay for avian influenza A (H5N1) virus. 3,3´,5,5´- Tetramethylbenzidine (TMBZ) was used as a reducing agent to produce gold nanoparticles (Au NPs) with blue colored solution from a viral target-specific antibody-gold ion mixture at first step. The developed blue color from the sensing design was further amplified through catalytic activity of Au NPs in presence of TMBZ-hydrogen peroxide (H2O2) solution in second step. Hence, the developed dual enhanced colorimetric immunosensor enables the detection of avian influenza virus A (H5N1) with a limit of detection (LOD) of 1.11 pg/mL. Our results confirmed that the developed assay has superior sensitivity than the conventional ELISA method, plasmonic-based bioassay and commercial flu diagnostic kits. Proposed sensing method further showed its capability to detect viruses, avian influenza A (H4N6) and A (H9N2) virus, in blood samples with limit of detection of 0.0269 HAU and 0.0331 HAU respectively.

15.
Sensors (Basel) ; 17(5)2017 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-28489036

RESUMO

Enzyme-linked immunosorbent assay (ELISA) is a popular assay technique for the detection and quantification of various biological substances due its high sensitivity and specificity. More often, it requires large and expensive laboratory instruments, which makes it difficult to conduct when the tests must be performed quickly at the point-of-care (POC). To increase portability and ease of use, we propose a portable diagnostic system based on a Raspberry Pi imaging sensor for the rapid detection of progesterone in milk samples. We designed, assembled, and tested a standalone portable diagnostic reader and validated it for progesterone detection against a standard ELISA assay using a commercial plate reader. The portable POC device yielded consistent results, regardless of differences in the cameras and flashlights between various smartphone devices. An Android application was built to provide front-end access to users, control the diagnostic reader, and display and store the progesterone measurement on the smartphone. The diagnostic reader takes images of the samples, reads the pixel values, processes the results, and presents the results on the handheld device. The proposed POC reader can perform to superior levels of performance as a plate reader, while adding the desirable qualities of portability and ease of use.


Assuntos
Smartphone , Animais , Colorimetria , Ensaio de Imunoadsorção Enzimática , Leite , Progesterona
16.
Sci Rep ; 7: 44495, 2017 03 14.
Artigo em Inglês | MEDLINE | ID: mdl-28290527

RESUMO

Nanomaterials without chemical linkers or physical interactions that reside on a two-dimensional surface are attractive because of their electronic, optical and catalytic properties. An in situ method has been developed to fabricate gold nanoparticle (Au NP) films on different substrates, regardless of whether they are hydrophilic or hydrophobic surfaces, including glass, 96-well polystyrene plates, and polydimethylsiloxane (PDMS). A mixture of sodium formate (HCOONa) and chloroauric acid (HAuCl4) solution was used to prepare Au NP films at room temperature. An experimental study of the mechanism revealed that film formation is dependent on surface wettability and inter particle attraction. The as-fabricated Au NP films were further applied to the colorimetric detection of influenza virus. The response to the commercial target, New Caledonia/H1N1/1999 influenza virus, was linear in the range from 10 pg/ml to 10 µg/ml and limit of detection was 50.5 pg/ml. In the presence of clinically isolated influenza A virus (H3N2), the optical density of developed color was dependent on the virus concentration (10-50,000 PFU/ml). The limit of detection of this study was 24.3 PFU/ml, a limit 116 times lower than that of conventional ELISA (2824.3 PFU/ml). The sensitivity was also 500 times greater than that of commercial immunochromatography kits.


Assuntos
Técnicas Biossensoriais , Vírus da Influenza A Subtipo H1N1/isolamento & purificação , Vírus da Influenza A Subtipo H3N2/isolamento & purificação , Influenza Humana/diagnóstico , Ouro/química , Humanos , Interações Hidrofóbicas e Hidrofílicas , Vírus da Influenza A Subtipo H1N1/patogenicidade , Vírus da Influenza A Subtipo H3N2/patogenicidade , Influenza Humana/virologia , Nanopartículas Metálicas/química
17.
Biosens Bioelectron ; 87: 558-565, 2017 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-27611475

RESUMO

A hybrid structure of graphene-gold nanoparticles (Grp-Au NPs) was designed as a new nanoprobe for colorimetric immunoassays. This hybrid structure was prepared using chloroauric acid, sodium formate and Grp flakes at room temperature. Au NPs attached strongly onto the Grp surface, and their size was controlled by varying the sodium formate concentration. The Raman intensity of the Grp-Au NP hybrids was significantly enhanced at 1567cm-1 and 2730cm-1 compared with those of pristine Grp because of the electronic interaction between Au NPs and Grp. The Grp-Au NPs with a hybrid structure catalyzed the oxidation of the peroxidase substrate 3,3,5,5-tetramethylbenzidine (TMB) with H2O2, developing a blue color in aqueous solution. This catalytic activity was utilized to detect norovirus-like particles (NoV-LPs) in human serum. The enhanced colorimetric response was monitored using Ab-conjugated-Grp-Au NPs and found to depend on the NoV-LP concentration, exhibiting a linear response from 100pg/mL to 10µg/mL. The limit of detection (LOD) of this proposed method was 92.7pg/mL, 112 times lower than that of a conventional enzyme-linked immunosorbent assay (ELISA). The sensitivity of this test was also 41 times greater than that of a commercial diagnostic kit. The selectivity of the Grp-Au NPs was tested with other viruses, and no color changes were observed. Therefore, the proposed system will facilitate the utilization of the intrinsic peroxidase-like activity of Grp-Au NPs in medical diagnostics. We believe that the engineered catalytic Grp-Au NP hybrids could find potential applications in the future development of biocatalysts and bioassays.


Assuntos
Infecções por Caliciviridae/diagnóstico , Colorimetria/métodos , Ouro/química , Grafite/química , Nanopartículas/química , Norovirus/isolamento & purificação , Técnicas Biossensoriais/métodos , Infecções por Caliciviridae/sangue , Infecções por Caliciviridae/virologia , Humanos , Limite de Detecção , Nanopartículas/ultraestrutura , Peroxidase/química
18.
Biosens Bioelectron ; 85: 503-508, 2016 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-27209577

RESUMO

Multifunctional nanohybrids have created new and valuable opportunities for a wide range of catalysis and biotechnology applications. Here, we present a relatively simple method for producing nanohybrids composed of gold nanoparticles (Au NPs) and carbon nanotubes (CNTs) that does not require an acidic pre-treatment of the CNTs. Transmission electron microscopy (TEM) images and ultraviolet-visible (UV-vis) spectra revealed that Au NPs bonded to the CNT surface. Surface-enhanced Raman scattering (SERS) revealed a stronger signal from Au-CNT nanohybrids than from pristine CNTs. The Au-CNT nanohybrids showed catalytic activity in the oxidation of 3, 3', 5, 5'-tetramethyl-benzidine (TMB) by H2O2 and developed a unique blue colour in aqueous solution. Because of the enhanced peroxidase-like activity of these Au-CNT nanohybrids, they were selected for use as part of a highly sensitive colorimetric test for influenza virus A (H3N2). In the presence of influenza A virus (H3N2) in the test system (specific antibody-conjugated Au CNT nanohybrids-TMB-H2O2), a deep blue colour developed, the optical density of which was dependent on the virus concentration (10-50,000 PFU/ml). The limit of detection of this proposed method was 3.4 PFU/ml, a limit 385 times lower than that of conventional ELISA (1312 PFU/ml). The sensitivity of this test was also 500 times greater than that of commercial immunochromatography kits. The nanohybrid preparation and colorimetric detection methods reported herein may be easily adapted to other nanohybrid structures with enzyme mimetic properties for broader applications in catalysis and nanobiotechnology.


Assuntos
Técnicas Biossensoriais/métodos , Ouro/química , Vírus da Influenza A Subtipo H3N2/isolamento & purificação , Influenza Humana/sangue , Nanopartículas Metálicas/química , Nanotubos de Carbono/química , Anticorpos Imobilizados/química , Benzidinas/química , Catálise , Colorimetria/métodos , Humanos , Peróxido de Hidrogênio/química , Influenza Humana/diagnóstico , Limite de Detecção , Nanopartículas Metálicas/ultraestrutura , Nanotubos de Carbono/ultraestrutura , Peroxidase/química , Análise Espectral Raman/métodos
19.
Biotechnol Bioeng ; 113(10): 2298-303, 2016 10.
Artigo em Inglês | MEDLINE | ID: mdl-27002303

RESUMO

A modified enzyme-linked immunosorbent assay (ELISA) with nanomaterials is an effective and powerful method to amplify the signal and reduce the cost of detecting and measuring trace biomarkers or proteins. In this study, an ultra-sensitive colorimetric immunoassay was designed, and its ability to detect influenza viruses using positively charged gold nanoparticles ((+)Au NPs) was assessed as a possible role for peroxidase-mimic inorganic enzymes. This method detected influenza virus A (H1N1) with a linear range up to 10 pg mL(-1) and clinically isolated influenza virus A (H3N2) up to 10 plaque forming units (PFU) mL(-1) , where its sensitivity improved to 500-fold higher than that of commercial virus kits. The sensitivity of this proposed method was not declined even though in complex biological media in compared to conventional ELISA. These results revealed that the (+)AuNP-based colorimetric immunoassay could be suitable for lab-on-a-chip device and open new opportunities for clinical protein diagnostics. Biotechnol. Bioeng. 2016;113: 2298-2303. © 2016 Wiley Periodicals, Inc.


Assuntos
Colorimetria/instrumentação , Ouro/química , Peróxido de Hidrogênio/química , Vírus da Influenza A Subtipo H3N2/isolamento & purificação , Nanopartículas Metálicas/química , Peroxidase/química , Benzidinas/química , Técnicas Biossensoriais/instrumentação , Desenho de Equipamento , Análise de Falha de Equipamento , Vírus da Influenza A Subtipo H3N2/imunologia , Peroxidase/imunologia , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
20.
Nanoscale Res Lett ; 11(1): 65, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26847691

RESUMO

The demand for biologically compatible and stable noble metal nanoparticles (NPs) has increased in recent years due to their inert nature and unique optical properties. In this article, we present 11 different synthetic methods for obtaining gold nanoparticles (Au NPs) through the use of common biological buffers. The results demonstrate that the sizes, shapes, and monodispersity of the NPs could be varied depending on the type of buffer used, as these buffers acted as both a reducing agent and a stabilizer in each synthesis. Theoretical simulations and electrochemical experiments were performed to understand the buffer-dependent variations of size and morphology exhibited by these Au NPs, which revealed that surface interactions and the electrostatic energy on the (111) surface of Au were the determining factors. The long-term stability of the synthesized NPs in buffer solution was also investigated. Most NPs synthesized using buffers showed a uniquely wide range of pH stability and excellent cell viability without the need for further modifications.

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