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1.
Analyst ; 148(19): 4857-4868, 2023 Sep 25.
Artigo em Inglês | MEDLINE | ID: mdl-37624366

RESUMO

Electrochemical sensing is ubiquitous in a number of fields ranging from biosensing, to environmental monitoring through to food safety and battery or corrosion characterisation. Whereas conventional potentiostats are ideal to develop assays in laboratory settings, they are in general, not well-suited for field work due to their size and power requirements. To address this need, a number of portable battery-operated potentiostats have been proposed over the years. However, most open source solutions do not take full advantage of integrated circuit (IC) potentiostats, a rapidly evolving field. This is partly due to the constraining requirements inherent to the development of dedicated interfaces, such as apps, to address and control a set of common electrochemical sensing parameters. Here we propose the PocketEC, a universal app that has all the functionalities to interface with potentiostat ICs through a user defined property file. The versatility of PocketEC, developed with an assay developer mindset, was demonstrated by interfacing it, via Bluetooth, to the ADuCM355 evaluation board, the open-source DStat potentiostat and the Voyager board, a custom-built, small footprint potentiostat based around the LMP91000 chip. The Voyager board is presented here for the first time. Data obtained using a standard redox probe, Ferrocene Carboxylic Acid (FCA) and a silver ion assay using anodic stripping multi-step amperometry were in good agreement with analogous measurements using a bench top potentiostat. Combined with its Voyager board companion, the PocketEC app can be used directly for a number of wearable or portable electrochemical sensing applications. Importantly, the versatility of the app makes it a candidate of choice for the development of future portable potentiostats. Finally, the app is available to download on the Google Play store and the source codes and design files for the PocketEC app and the Voyager board are shared via Creative Commons license (CC BY-NC 3.0) to promote the development of novel portable or wearable applications based on electrochemical sensing.

2.
Analyst ; 147(21): 4866-4875, 2022 Oct 24.
Artigo em Inglês | MEDLINE | ID: mdl-36226569

RESUMO

Herein, a unique electrochemiluminescence (ECL) sensor combined with a paper electrode was proposed for the detection of phenylalanine (L-Phe) in blood samples. The L-Phe detection was performed by converting L-Phe into ammonia using phenylalanine ammonia-lyase (PAL) enzyme and the ECL signal of Ru(bpy)32+ was produced in combination with ammonia as a co-reactant. In this report, we used AuNP decorated paper electrodes to obtain the ECL signal of Ru-(bpy)32+ in the presence of NH3. The produced ammonia molecules were carried to the working electrode and their contact with the sample was cut off. Since there was no contact between the sample matrix and the electrode, the developed method achieved excellent selectivity. According to experimental data, a linear increase of the ECL signals with the logarithms of varying L-Phe concentrations between 1.5 × 10-2 and 1.211 mM was observed with a correlation coefficient (R2) of 0.9898 and a limit of detection (LOD) of 8.4 × 10-3 mM. The proposed method was efficiently applied for L-Phe detection in reference blood samples and the average recovery was calculated as 96.8%. Furthermore, the HPLC-UV method as a comparison verified that the recovery values provided by the proposed method were acceptable with a similarity of 95.1% for L-Phe detection in a reference blood sample. The results showed that the developed ECL sensor combined with the paper electrode can be considered as a promising unique and powerful technique for a point-of-care diagnosis of PKU.


Assuntos
Técnicas Biossensoriais , Técnicas Eletroquímicas , Técnicas Eletroquímicas/métodos , Medições Luminescentes/métodos , Amônia , Fenilalanina , Fenilalanina Amônia-Liase , Eletrodos , Técnicas Biossensoriais/métodos
3.
J Dairy Sci ; 105(4): 2895-2907, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-35181133

RESUMO

Salmonella, as a common foodborne pathogen in dairy products, poses a great threat to human health. We studied a new detection method based on quantum dots (QD). A fluorescent biosensor with multiple fluorescent signal amplification based on a streptavidin (SA) biotin system and the polyamino linear polymer poly-l-lysine (PLL) were established to detect Salmonella in milk. First, Salmonella was captured on a black 96-well plate with paired Salmonella mAb to form a double-antibody sandwich. Second, SA was immobilized on biotin-modified mAb by SA-biotin specific bond. Then, the biotin-modified polylysine (BT-PLL) was bound on SA and specifically bonded again through the SA-biotin system. Finally, water-soluble CdSe/ZnS QD-labeled SA was added to a black 96-well plate for covalent coupling with BT-PLL. The fluorescent signal was amplified in a dendritic manner by the layer-by-layer overlap of SA and biotin and the covalent coupling of biotinylated PLL. Under optimal conditions, the detection limit was 4.9 × 103 cfu/mL in PBS. The detection limit was 10 times better than that of the conventional sandwich ELISA. In addition, the proposed biosensor was well specific and could be used for detecting Salmonella in milk samples.


Assuntos
Técnicas Biossensoriais , Pontos Quânticos , Animais , Técnicas Biossensoriais/veterinária , Biotina/química , Leite , Polilisina , Salmonella , Estreptavidina/química
4.
Pharm Dev Technol ; 27(7): 794-804, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-36046958

RESUMO

The choice of polymer and its compatibility with drug used determine the fate of nanoparticle in therapy. There has been limited sources about effect of resomer differentiation in nanoparticle related with physical and chemical properties and also biological activities of product. Therefore, we aimed to formulate docetaxel-loaded polylactic-co-glycolic acid nanoparticles with different molecular weights (Resomer 502 and 504) and terminal groups (Resomer 502H and 504H) and to investigate the effect of these resomers on nanoparticle character, prostate cancer, and healthy cells. Docetaxel-loaded PLGA nanoparticles were prepared by single emulsion solvent evaporation method. Surface characterizations were carried out by zeta sizer and scanning electron microscopy. Encapsulation efficiency, in vitro drug release profiles, and cytotoxic activity were determined. Main effect on the surface morphology of nanoparticles was the molecular weight of the polymer. The groups with acid terminal function have higher encapsulation and reaction efficiency. In all formulations, in vitro release was observed after 334 h at pH 7.4 and 240 h at pH 5.6. Also, the groups with high molecular weight showed selective cytotoxicity. These resomers especially RG 504 and RG 504H have potential to be used as a low-dose and high-efficiency extended-release drug delivery system in the treatment of prostate cancer.


Assuntos
Antineoplásicos , Nanopartículas , Neoplasias de Próstata Resistentes à Castração , Antineoplásicos/química , Antineoplásicos/farmacologia , Docetaxel/química , Portadores de Fármacos/química , Emulsões , Humanos , Masculino , Peso Molecular , Nanopartículas/química , Tamanho da Partícula , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/química , Polímeros/química , Solventes
5.
Turk J Med Sci ; 50(5): 1470-1479, 2020 08 26.
Artigo em Inglês | MEDLINE | ID: mdl-32178510

RESUMO

Background: Heroin can be detected and quantified by certain analytical methods, however, forensic professionals and criminal laboratories study for cheaper and faster detection tools. Surface-enhanced Raman spectroscopy (SERS) rises as a possible alternative tool with its widening application spectra. There are few studies regarding Raman and SERS spectra of heroin and its metabolites, which are unfortunately controversial. In this study, we compared five different surfaces in order to find out more efficient Raman-active substrate for opiate detection and rapid quantification of heroin and its metabolites in saliva. Materials and methods: Morphine standard material was used to identify proper surface for SERS analysis of opiates. Heroin and its metabolites (morphine, morphine-3-ß-glucuronide and 6-monoacetyl morphine) were calibrated between 50 ppb and 500 ppm and quantified on AuNRs with signal enhancement of silver colloids in saliva. Raman microscope with a 785-nm laser source was used. Results and Conclusion: Obtained results showed that heroin and its metabolites can be detected and quantified in saliva samples using a SERS-based system. Additionally, the present study revealed that synergetic effect of a specific gold nano-surface with ability controlling liquid motion and silver nanoparticles increase band numbers and intensities. Therefore, we suggest a fast, accurate and cost-effective method to detect and quantify heroin in biological fluids


Assuntos
Heroína/análise , Saliva/química , Análise Espectral Raman/métodos , Adulto , Ouro/química , Humanos , Nanopartículas Metálicas/química , Morfina/análise , Prata/química
6.
Turk J Med Sci ; 50(4): 1157-1167, 2020 06 23.
Artigo em Inglês | MEDLINE | ID: mdl-32283902

RESUMO

Background/aim: We aimed to develop a rapid method to enumerate Listeria monocytogenes (L. monocytogenes) utilizing magnetic nanoparticle based preconcentration and surface-enhanced Raman spectroscopy measurements. Materials and methods: Biological activities of magnetic Au-nanoparticles have been observed to have the high biocompatibility, and a sample immunosensor model has been designed to use avidin attached Au-nanoparticles for L. monocytogenes detection. Staphylococcus aureus (S. aureus) and Salmonella typhimurium (S. typhimurium) bacteria cultures were chosen for control studies. Antimicrobial activity studies have been done to identify bio-compatibility and bio-characterization of the Au-nanoparticles in our previous study and capturing efficiencies to bacterial surfaces have been also investigated. Results: We constructed the calibration graphs in various population density of L. monocytogenes as 2.2 × 101 to 2.2 × 106 cfu/mL and the capture efficiency was found to be 75%. After the optimization procedures, population density of L. monocytogenes and Raman signal intensity showed a good linear correlation (R2 = 0.991) between 102 to 106 cfu/mL L. monocytogenes. The presented sandwich assay provides low detection limits and limit of quantification as 12 cfu/mL and 37 cfu/mL, respectively. We also compared the experimental results with reference plate-counting methods and the practical utility of the proposed assay is demonstrated using milk samples. Conclusion: It is focused on the enumeration of L. monocytogenes in milk samples and the comparision of results of milk analysis obtained by the proposed SERS method and by plate counting method stay in food agreement. In the present study, all parameters were optimized to select SERS-based immunoassay method for L. monocytogenes bacteria to ensure LOD, selectivity, precision and repeatablity.


Assuntos
Separação Imunomagnética/métodos , Listeria monocytogenes/imunologia , Leite/microbiologia , Análise Espectral Raman/métodos , Animais , Anticorpos Antibacterianos/análise , Materiais Biocompatíveis , Qualidade de Produtos para o Consumidor , Contaminação de Alimentos/análise , Microbiologia de Alimentos , Ouro , Magnetismo , Nanopartículas Metálicas , Salmonella typhimurium , Sensibilidade e Especificidade , Staphylococcus aureus
7.
Analyst ; 144(11): 3573-3580, 2019 Jun 07.
Artigo em Inglês | MEDLINE | ID: mdl-31062777

RESUMO

Beta-hemolytic, Group A Streptococcus pyogenes (GAS) is a life-threating pathogen and the reason for prominent disease, pharyngitis. The conventional analysis of GAS, gold standard, takes 48 hours and the related rapid tests lack in accuracy and sensitivity. In this study, firstly, the efficiency of swab sampling, which is a must in the GAS detection, was discussed with the proposed surface-enhanced Raman spectroscopy (SERS)-based batch assay and each step was controlled by the plate-counting method. Secondly, SERS-based lateral flow immunoassay (LFIA) test strips were constructed and the variation in the SERS intensity of 5,5'-dithiobis (2-nitrobenzoic acid) (DTNB) was observed. Thus, a linear correlation was found with a R2 value of 0.9926 and the LOD was calculated to be 0.2 CFU mL-1 of GAS which could be counted as one cell. The combination of the gold standard with the LFIA-SERS technique enabled the fast and accurate pathogen detection. In addition, GAS was quantified with paper-based test strips up to 100 CFU ml-1 level of bacteria for the first time without any interference. Besides, this study was featured with the discussion of the whole cell and pretreated cell detection of pathogens with LFIAs. Therefore, this work enlightens the points that have never been discussed on pathogen detection with paper-based platforms.


Assuntos
Streptococcus pyogenes/isolamento & purificação , Anticorpos/imunologia , Ácido Ditionitrobenzoico/química , Ouro/química , Imunoensaio/métodos , Limite de Detecção , Nanopartículas Metálicas/química , Saliva/microbiologia , Análise Espectral Raman/métodos , Streptococcus pyogenes/imunologia
8.
Nat Mater ; 16(9): 918-924, 2017 09.
Artigo em Inglês | MEDLINE | ID: mdl-28783157

RESUMO

π-Conjugated organic semiconductors have been explored in several optoelectronic devices, yet their use in molecular detection as surface-enhanced Raman spectroscopy (SERS)-active platforms is unknown. Herein, we demonstrate that SERS-active, superhydrophobic and ivy-like nanostructured films of a molecular semiconductor, α,ω-diperfluorohexylquaterthiophene (DFH-4T), can be easily fabricated by vapour deposition. DFH-4T films without any additional plasmonic layer exhibit unprecedented Raman signal enhancements up to 3.4 × 103 for the probe molecule methylene blue. The combination of quantum mechanical computations, comparative experiments with a fluorocarbon-free α,ω-dihexylquaterthiophene (DH-4T), and thin-film microstructural analysis demonstrates the fundamental roles of the π-conjugated core fluorocarbon substitution and the unique DFH-4T film morphology governing the SERS response. Furthermore, Raman signal enhancements up to ∼1010 and sub-zeptomole (<10-21 mole) analyte detection were accomplished by coating the DFH-4T films with a thin gold layer. Our results offer important guidance for the molecular design of SERS-active organic semiconductors and easily fabricable SERS platforms for ultrasensitive trace analysis.


Assuntos
Materiais Revestidos Biocompatíveis/química , Fluorocarbonos/química , Ouro/química , Membranas Artificiais , Semicondutores , Tiofenos/química , Análise Espectral Raman
9.
Anal Bioanal Chem ; 408(29): 8447-8456, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27722945

RESUMO

We present a surface-enhanced Raman probe (SERS) platform for the determination of a prohibited substance, recombinant erythropoietin (rEPO), in urine matrix, using nanoparticles as substrate. Rod-shaped gold nanoparticles (GNR) were modified with a Raman label and an antibody as SERS probe. We developed two SERS-based immunoassays for detection and quantification of rEPO in urine. In the first assay, rEPO was determined by a sandwich assay with gold surfaces and GNR. In the second assay, rEPO was extracted by using core shell-structured magnetic iron oxide gold nanoparticles, and again sandwich assay was performed by using GNR. We also demonstrated the ability of the proposed method to discriminate rEPO and urinary erythropoietin (uEPO). A good linear correlation was obtained between logarithms of rEPO concentrations in urine and Raman intensities within the range of 10-1-103 pg mL-1 rEPO concentrations. Detection limits which are smaller than 0.1 pg mL-1 levels were achieved owing to the high extractive performance of the nanoextraction techniques. Graphical Abstract Schematic represantation of surface-enhanced Raman probe for rapid nanoextraction and detection of erythropoietin.


Assuntos
Eritropoetina/urina , Ouro/química , Nanopartículas de Magnetita/química , Substâncias para Melhoria do Desempenho/urina , Análise Espectral Raman/métodos , Anticorpos Imobilizados/química , Dopagem Esportivo/legislação & jurisprudência , Humanos , Limite de Detecção , Proteínas Recombinantes/urina , Propriedades de Superfície
10.
Toxicol Mech Methods ; 26(6): 414-8, 2016 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-27278718

RESUMO

The popularity of electronic cigarettes (e-cigarettes) is rapidly increasing in many countries. These devices are designed to imitate regular cigarettes, delivering nicotine via inhalation without combusting tobacco but currently, there is a lack of scientific evidence on the presence or absence of nicotine exposure. Such research relies on evidence from e-cigarette users urine samples. In this study, we aimed to determine the levels and compare the amount of nicotine to which e-cigarette users, cigarette smokers and passive smokers are exposed. Therefore, urine samples were collected from e-cigarette users, cigarette smokers, passive smokers, and healthy nonsmokers. The urinary cotinine levels of the subjects were determined using gas chromatography-mass spectrometry. The mean (±SD) urinary cotinine levels were determined as 1755 ± 1848 ng/g creatinine for 32 e-cigarette users, 1720 ± 1335 ng/g creatinine for 33 cigarette smokers and 81.42 ± 97.90 ng/g creatinine for 33 passive smokers. A significant difference has been found between cotinine levels of e-cigarette users and passive smokers (p < 0.05). There were no statistically significant differences between e-cigarette users and cigarette smokers (p > 0.05). This is a seminal study to demonstrate the e-cigarette users are exposed to nicotine as much as cigarette smokers.


Assuntos
Cotinina/urina , Creatinina/urina , Sistemas Eletrônicos de Liberação de Nicotina/efeitos adversos , Nicotina , Fumar/urina , Poluição por Fumaça de Tabaco/análise , Adulto , Biomarcadores/urina , Estudos de Casos e Controles , Feminino , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Masculino , Nicotina/administração & dosagem , Nicotina/farmacocinética , Fumar/efeitos adversos , Inquéritos e Questionários
11.
Anal Bioanal Chem ; 407(27): 8243-51, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-26363778

RESUMO

In this report, we present a paper membrane-based surface-enhanced Raman scattering (SERS) platform for the determination of blood glucose level using a nitrocellulose membrane as substrate paper, and the microfluidic channel was simply constructed by wax-printing method. The rod-shaped gold nanorod particles were modified with 4-mercaptophenylboronic acid (4-MBA) and 1-decanethiol (1-DT) molecules and used as embedded SERS probe for paper-based microfluidics. The SERS measurement area was simply constructed by dropping gold nanoparticles on nitrocellulose membrane, and the blood sample was dropped on the membrane hydrophilic channel. While the blood cells and proteins were held on nitrocellulose membrane, glucose molecules were moved through the channel toward the SERS measurement area. Scanning electron microscopy (SEM) was used to confirm the effective separation of blood matrix, and total analysis is completed in 5 min. In SERS measurements, the intensity of the band at 1070 cm(-1) which is attributed to B-OH vibration decreased depending on the rise in glucose concentration in the blood sample. The glucose concentration was found to be 5.43 ± 0.51 mM in the reference blood sample by using a calibration equation, and the certified value for glucose was 6.17 ± 0.11 mM. The recovery of the glucose in the reference blood sample was about 88 %. According to these results, the developed paper-based microfluidic SERS platform has been found to be suitable for use for the detection of glucose in blood samples without any pretreatment procedure. We believe that paper-based microfluidic systems may provide a wide field of usage for paper-based applications.


Assuntos
Glicemia/análise , Colódio/química , Membranas Artificiais , Técnicas Analíticas Microfluídicas/instrumentação , Papel , Análise Espectral Raman/instrumentação , Desenho de Equipamento , Ouro/química , Limite de Detecção , Nanotubos/química
12.
Analyst ; 139(5): 1141-7, 2014 Mar 07.
Artigo em Inglês | MEDLINE | ID: mdl-24418951

RESUMO

In this study, two different assay methods were developed using a surface enhanced Raman scattering (SERS) label for sensitive miR-21 detection. In the first method (direct assay), the miR-21 probe was attached to SERS-labelled, rod-shaped gold nanoparticles and hybridised with the target miR-21, which was previously immobilised onto the gold slide. In the second method (sandwich assay), the target miR-21 was captured by an miR-21 probe immobilised onto the gold slide and hybridised with a second miR-21 probe immobilised on the SERS-labeled, rod-shaped gold nanoparticles. SERS signals of developed assays were obtained via a SERS spectrum of 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) on the rod-shaped nanoparticles. The calibration curves were plotted to measure the different concentrations of miR-21. The detection limits of the direct and sandwich assays, which last less than 40 min, were found to be 0.36 and 0.85 nM, respectively. The developed SERS-based methods offer rapid, selective, sensitive and easy detection of miR-21, especially compared to conventional PCR-based methods.


Assuntos
Técnicas Biossensoriais/métodos , Ouro/química , Nanopartículas Metálicas/química , MicroRNAs/análise , Análise Espectral Raman/métodos , Oligonucleotídeos/análise
13.
Phys Chem Chem Phys ; 16(12): 5563-70, 2014 Mar 28.
Artigo em Inglês | MEDLINE | ID: mdl-24514029

RESUMO

The detection of molecules at an ultralow level by Surface-Enhanced Raman Spectroscopy (SERS) has recently attracted enormous interest for various applications especially in biological, medical, and environmental fields. Despite the significant progress, SERS systems are still facing challenges for practical applications related to their sensitivity, reliability, and selectivity. To overcome these limitations, in this study, we have proposed a simple yet facile concept by combining 3-D anisotropic gold nanorod arrays with colloidal gold nanoparticles having different shapes for highly reliable, selective, and sensitive detection of some hazardous chemical and biological warfare agents in trace amounts through SERS. The gold nanorod arrays were created on the BK7 glass slides or silicon wafer surfaces via the oblique angle deposition (OAD) technique without using any template material or lithography technique and their surface densities were adjusted by manipulating the deposition angle (α). It is found that gold nanorod arrays fabricated at α = 10° exhibited the highest SERS enhancement in the absence of colloidal gold nanoparticles. Synergetic enhancement was obviously observed in SERS signals when combining gold nanorod arrays with colloidal gold nanoparticles having different shapes (i.e., spherical, rod, and cage). Due to their ability to produce localized surface plasmons (LSPs) in transverse and longitudinal directions, utilization of colloidal gold nanorods as a synergetic agent led to an increase in the enhancement factor by about tenfold compared to plain gold nanorod arrays. Moreover, we have tested our approach to detect some chemical and biological toxins namely dipicolinic acid (DIP), methyl parathion (MP), and diethyl phosphoramidate (DP). For all toxins, Raman spectra with high signal-to-noise ratios and reproducibility were successfully obtained over a broad concentration range (5 ppm-10 ppb). Our results suggest that the slightly tangled and closely-packed anisotropic gold nanorod arrays reinforced by the gold nanoparticles may serve as an ideal SERS substrate to detect any analyte in trace amounts.


Assuntos
Ouro/química , Nanopartículas Metálicas/química , Nanotubos/química , Amidas/análise , Coloides/química , Metil Paration/análise , Compostos Organofosforados/análise , Tamanho da Partícula , Ácidos Picolínicos/análise , Análise Espectral Raman , Propriedades de Superfície
14.
Forensic Sci Int ; 354: 111885, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-38007869

RESUMO

Various body fluids such as blood, semen, vaginal secretions, and saliva are frequently encountered at crime scene. In cases of sexual assault, semen stains are one of the most reliable evidence of biological origin. In this study, our objective was to develop a method for estimating the time since deposition of semen stains on five different fabric types using Attenuated Total Reflectance-Fourier Transform Infrared (ATR-FTIR) Spectroscopy, with a focus on a time frame of up to 8 weeks. Semen samples from six different volunteers were dripped onto five distinct fabric materials, and ATR-FTIR measurements were obtained at 17 different time points. Principal component analysis (PCA) and partial least squares (PLS) methods were employed to differentiate semen stains on various fabric samples and estimate the age of semen stains. Models constructed using PCA and PLSR achieved high R2 values and low root-mean-square error (RMSE). While the performance varies depending on fabric types, it was observed that age estimation of semen stains can be made within following intervals: 0.39-0.76 days for 0-7 day range, 2.59-3.38 days for the 1-8 week range, and 3.98-8.1 days for the 0-56 day range. This study demonstrates the effectiveness of using ATR-FTIR spectroscopy in combination with chemometrics to estimate the age of human semen stains on various fabric types based on time-dependent spectral changes.


Assuntos
Líquidos Corporais , Sêmen , Feminino , Humanos , Recém-Nascido , Sêmen/química , Espectroscopia de Infravermelho com Transformada de Fourier/métodos , Corantes/análise , Quimiometria , Líquidos Corporais/química , Análise dos Mínimos Quadrados , Proteínas Mutadas de Ataxia Telangiectasia/análise
15.
Turk J Chem ; 48(4): 676-690, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39296784

RESUMO

The analysis of substances and samples obtained from a crime scene is very important in solving forensic cases. To determine the variables involved in a crime and to expedite the investigation process, the rapid analysis of body fluids in small quantities and within environments containing diverse components is particularly necessary. For this reason, it is of great importance to analyze biological fluids with rapid, noncontaminating, nondestructive, low-cost, and accurate techniques. In recent years, with advancements in laser technology, spectroscopic methods have been introduced as analytical techniques in forensic medicine and chemical studies. This study focuses on surface-enhanced Raman spectroscopy (SERS) to demonstrate the detection of blood samples in simulated crime scenes. To minimize the background signal from fluorescent biomolecules in blood, dilution was performed with two different components and Raman analysis was performed for four different concentrations of blood. In general, a decrease in noise in the spectra was observed as the blood was diluted. Crime scenes consisting of pure blood, blood diluted with ethanol and distilled water (1:2, 1:4, and 1:8), a blood-mineral water mixture, a blood-cherry juice mixture, and silver nanoparticle-added mixtures were simulated, and their spectra were examined. Chemometric analyses of the data were performed. Despite high noise and low peak intensities, blood-identifying signals were detected when examining different blood concentrations. It was observed that silver nanoparticles provided high enhancement of blood peaks thanks to their strong plasmonic properties.

16.
Bioelectrochemistry ; 160: 108785, 2024 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-39094446

RESUMO

This study presents a simple, fast, and sensitive label-free sensing assay for the precise enumeration of modeled pathogenic Escherichia coli K12 (E. coli K12) bacteria for the first time. The method employs the covalent binding bacteriophage technique on the surface of a reversible addition-fragmentation chain transfer (RAFT) polymer film. The Nyquist plots obtained from electrochemical impedance spectroscopy (EIS) identified the charge transfer resistance Rct was calculated from a suitable electrochemical circuit model through an evaluation of the relevant parameter after the immobilization of the bacteriophage and the binding of specific E. coli K12. The impedimetric biosensor reveals specific and reproducible detection with sensitivity in the linear working range of 104.2-107.0 CFU/mL, a limit of detection (LOD) of 101.3 CFU/mL, and a short response time of 15 min. The SERS response validates the surface roughness and interaction of the SERS-tag with E. coli K12-modified electrodes. Furthermore, the covalently immobilized active phage selectivity was proved against various non-targeting bacterial strains in the presence of targeted E.coli K12 with a result of 94 % specificity and 98 % sensitivity. Therefore, the developed phage-based electrode surface can be used as a disposable, label-free impedimetric biosensor for rapid and real-time monitoring of serum samples.


Assuntos
Técnicas Biossensoriais , Escherichia coli K12 , Limite de Detecção , Polímeros , Escherichia coli K12/virologia , Escherichia coli K12/isolamento & purificação , Técnicas Biossensoriais/métodos , Polímeros/química , Espectroscopia Dielétrica , Bacteriófagos , Eletrodos , Propriedades de Superfície , Análise Espectral Raman/métodos
17.
Biomacromolecules ; 14(9): 3001-9, 2013 Sep 09.
Artigo em Inglês | MEDLINE | ID: mdl-23885927

RESUMO

In this study, a simple and highly selective homogeneous sandwich assay was developed for fast and ultrasensitive detection of the tau protein using a combination of monoclonal antitau functionalized hybrid magnetic nanoparticles and polyclonal antitau immobilized gold nanoparticles as the recognition and surface-enhanced Raman scattering (SERS) component, respectively. The magnetic silica particles were first coated with poly(2-hydroxyethyl methacrylate) via surface-mediated reversible addition-fragmentation chain transfer (RAFT) polymerization and then biofunctionalized with monoclonal antitau, which are both specific for tau and can be collected via a simple magnet. After separating tau from the sample matrix, they were sandwiched with the SERS substrate composed of polyclonal antitau and 5,5-dithiobis(2-dinitrobenzoic acid) on gold nanoparticles. The correlation between the tau concentration and SERS signal was found to be linear within the range of 25 fM to 500 nM. The limit of detection for the sandwich assay is less than 25 fM. Moreover, the sandwich assay was also evaluated for investigating the tau specificity on bovine serum albumin and immunoglobulin G.


Assuntos
Doença de Alzheimer/diagnóstico , Proteínas tau/química , Humanos , Imunoglobulina G/química , Separação Imunomagnética , Limite de Detecção , Nanopartículas de Magnetita/química , Tamanho da Partícula , Polimerização , Soroalbumina Bovina/química , Espectroscopia de Infravermelho com Transformada de Fourier , Análise Espectral Raman/métodos
18.
Analyst ; 138(23): 7238-45, 2013 Dec 07.
Artigo em Inglês | MEDLINE | ID: mdl-24133677

RESUMO

A general protocol to prepare surface molecularly imprinted polymer core-shell superparamagnetic Fe3O4 nanoparticles (Fe3O4@MIP SPNPs), using a surface-mediated RAFT polymerization approach, is described. Cholesterol-imprinted Fe3O4@MIP SPNPs were obtained by oleic acid-stabilized Fe3O4 nanoparticles with a trithiocarbonate agent and subsequently by polymerizing thin molecularly imprinted layers composed of dimethylacrylamide and N,N'-methylene(bis)acrylamide units. The surface-mediated RAFT polymerization approach allows the synthesis of ∼20 nm hybrid composite particles with a ∼6 nm MIP shell, exhibiting superparamagnetic properties (saturation magnetization = 35.4 emu g(-1)) and specific molecular recognition of cholesterol. The Fe3O4@MIP SPNPs show the capability of rapid enriching and separating cholesterol (∼3.1% in weight) and are renewable and cyclically exploited due to their monodispersive and superparamagnetic features. Moreover, under optimal conditions, the Fe3O4@MIP SPNP recoveries of spiked human serum, milk, yolk and beef were 91.6%, 93.6%, 92.4% and 91.2%, respectively. Finally, the method of molecular imprinting on superparamagnetic particles can be extended to a wide range of applications for cell sorting, biomolecule enrichment and separation, and drug delivery.


Assuntos
Colesterol/química , Compostos Férricos/química , Nanopartículas Metálicas , Impressão Molecular , Colesterol/isolamento & purificação , Microscopia Eletrônica de Transmissão
19.
Int J Mol Sci ; 14(3): 6223-40, 2013 Mar 18.
Artigo em Inglês | MEDLINE | ID: mdl-23507756

RESUMO

We report the preparation and characterization of spherical core-shell structured Fe3O4-Au magnetic nanoparticles, modified with two component self-assembled monolayers (SAMs) consisting of 3-mercaptophenylboronic acid (3-MBA) and 1-decanethiol (1-DT). The rapid and room temperature synthesis of magnetic nanoparticles was achieved using the hydroxylamine reduction of HAuCl4 on the surface of ethylenediaminetetraacetic acid (EDTA)-immobilized iron (magnetite Fe3O4) nanoparticles in the presence of an aqueous solution of hexadecyltrimetylammonium bromide (CTAB) as a dispersant. The reduction of gold on the surface of Fe3O4 nanoparticles exhibits a uniform, highly stable, and narrow particle size distribution of Fe3O4-Au nanoparticles with an average diameter of 9 ± 2 nm. The saturation magnetization value for the resulting nanoparticles was found to be 15 emu/g at 298 K. Subsequent surface modification with SAMs against glucoside moieties on the surface of bacteria provided effective magnetic separation. Comparison of the bacteria capturing efficiency, by means of different molecular recognition agents 3-MBA, 1-DT and the mixed monolayer of 3-MBA and 1-DT was presented. The best capturing efficiency of E. coli was achieved with the mixed monolayer of 3-MBA and 1-DT-modified nanoparticles. Molecular specificity and selectivity were also demonstrated by comparing the surface-enhanced Raman scattering (SERS) spectrum of E. coli-nanoparticle conjugates with bacterial growth media.

20.
Mol Biotechnol ; 2023 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-37914863

RESUMO

Bacterial pathogens in water, food, and the environment are spreading diseases around the world. According to a World Health Organization (WHO) report, waterborne pathogens pose the most significant global health risks to living organisms, including humans and animals. Conventional bacterial detection approaches such as colony counting, microscopic analysis, biochemical analysis, and molecular analysis are expensive, time-consuming, less sensitive, and require a pre-enrichment step. However, the bacteriophage-based detection of pathogenic bacteria is a robust approach that utilizes bacteriophages, which are viruses that specifically target and infect bacteria, for rapid and accurate detection of targets. This review shed light on cutting-edge technologies about the novel structure of phages and the immobilization process on the surface of electrodes to detect targeted bacterial cells. Similarly, the purpose of this study was to provide a comprehensive assessment of bacteriophage-based biosensors utilized for pathogen detection, as well as their trends, outcomes, and problems. This review article summaries current phage-based pathogen detection strategies for the development of low-cost lab-on-chip (LOC) and point-of-care (POC) devices using electrochemical and optical methods such as surface-enhanced Raman spectroscopy (SERS).

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