RESUMO
In this study, an ultrasensitive detection platform for tobramycin (TOB) was developed, featuring a "sandwich" structure guided by AgNCs@PDANSs and Thi-AuNCs@ZnONSs. To address the issue of large background current peak signals in tagless sensors, Thi-AuNCs@ZnONSs composites were synthesized as signal tags. Zinc oxide nanosheets (ZnONSs) served as the loading agent, and AuNCs with the electroactive molecule Thi acted as carriers. Furthermore, AgNPs@PDANSs nanocomposites, possessing excellent electrical conductivity and large specific surface areas, were prepared as substrate materials for the modified electrodes. A "sandwich" structure strategy was also introduced to enhance the accuracy of the electrochemical aptasensor. This strategy, utilizing a dual sequence for target labeling and capture, yielded higher sensitivity and simplified the sensor construction compared to methods employing a single sequence. Under optimal conditions, the detection limit for TOB was established at 1.41 pM, with a detection range of 0.05-5000 nM. The aptasensor was effectively applied in the detection of TOB in tap and lake water, demonstrating outstanding reproducibility, selectivity, and stability. These results may serve as a reference for environmental TOB detection.
Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Técnicas Eletroquímicas , Nanopartículas Metálicas , Prata , Tobramicina , Poluentes Químicos da Água , Tobramicina/análise , Tobramicina/química , Aptâmeros de Nucleotídeos/química , Técnicas Eletroquímicas/métodos , Prata/química , Nanopartículas Metálicas/química , Poluentes Químicos da Água/análise , Técnicas Biossensoriais/métodos , Limite de Detecção , Antibacterianos/análise , Antibacterianos/química , Reprodutibilidade dos Testes , Óxido de Zinco/química , Lagos/análise , Lagos/químicaRESUMO
Using a chemiluminescence reaction between luminol and H2O2 in basic solution, an ultrasensitive electrochemiluminescence (ECL) aptasensor was developed for the determination of tobramycin (TOB), as an aminoglycoside antibiotic. Ti3C2/Ni/Sm-LDH-based nanocomposite effectively catalyzes the oxidation of luminol and decomposition of H2O2, leading to the formation of different reactive oxygen species (ROSs), thus amplifying the ECL signal intensity of luminol, which can be used for the determination of TOB concentration. To evaluate the performance of the electrochemiluminescence aptasensor and synthesized nanocomposite, different methods such as cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) analyses were performed. The considerable specific area, large number of active sites, and enhanced electron transfer reaction on this nanocomposite led to the development of an ECL aptasensor with high sensitivity and electrocatalytic activity. After optimizing the preparation method and analysis conditions, the aptasensor revealed a wide linear response ranging from 1.0 pM to 1.0 µM with a detection limit of 18 pM, displaying outstanding accuracy, specificity, and response stability. The developed ECL sensor was found to be applicable to the determination of TOB in human serum samples and is anticipated to possess excellent clinical potentials for detecting other antibiotics, as well.
Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Técnicas Eletroquímicas , Limite de Detecção , Medições Luminescentes , Nanocompostos , Tobramicina , Nanocompostos/química , Humanos , Técnicas Eletroquímicas/métodos , Aptâmeros de Nucleotídeos/química , Medições Luminescentes/métodos , Técnicas Biossensoriais/métodos , Tobramicina/sangue , Tobramicina/análise , Luminol/química , Antibacterianos/sangue , Antibacterianos/análise , Peróxido de Hidrogênio/química , Níquel/química , Titânio/químicaRESUMO
In this study, an ultrasensitive photoelectrochemical (PEC) aptasensor based on dual-sensitized heterojunction Ag2S/ZnS/NiS composites as a signal probe was proposed for the detection of tobramycin (TOB) by combining a cascaded quadratic signal amplification strategy. Specifically, compared to the limited visible light-harvesting capability of single sensitized composites, Ag2S/ZnS/NiS composites with p-n and n-n heterojunction could greatly improve the light energy utilization to tremendously strengthen the optical absorption in the entire visible-light region. Moreover, dual-sensitized heterojunction could effectively hinder the rapid recombination of photoelectrons and holes (carriers) to obtain a good photocurrent for improving the sensitivity of the aptasensor. Furthermore, a cascaded quadratic signal amplification strategy was applied to convert trace target TOB into plentiful gold nanoclusters (Au NCs) labelled double-stranded DNA for the construction of PEC aptasensor, with a broad linear detection range from 0.01 to 100 ng mL-1 and a low detection limit of 3.38 pg mL-1. Importantly, this study provided a versatile and sensitive PEC biosensing platform for TOB analysis, and demonstrated its successful application for TOB detection in milk samples. This protocol provides a novel dual-sensitized heterojunction composites to develop a highly efficient and harmfulless PEC aptasensor, which is expected to be used in food safety, environmental monitoring and other areas.
Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Técnicas Eletroquímicas , Luz , Limite de Detecção , Leite , Compostos de Prata , Sulfetos , Tobramicina , Compostos de Zinco , Tobramicina/análise , Tobramicina/química , Técnicas Eletroquímicas/métodos , Aptâmeros de Nucleotídeos/química , Compostos de Prata/química , Compostos de Zinco/química , Sulfetos/química , Leite/química , Animais , Nanopartículas Metálicas/química , Antibacterianos/análise , Ouro/química , Contaminação de Alimentos/análiseRESUMO
Electrochemical aptamer-based sensors support the high-frequency, real-time monitoring of molecules-of-interest in vivo. Achieving this requires methods for correcting the sensor drift seen during in vivo placements. While this correction ensures EAB sensor measurements remain accurate, as drift progresses it reduces the signal-to-noise ratio and precision. Here, we show that enzymatic cleavage of the sensor's target-recognizing DNA aptamer is a major source of this signal loss. To demonstrate this, we deployed a tobramycin-detecting EAB sensor analog fabricated with the DNase-resistant "xenonucleic acid" 2'O-methyl-RNA in a live rat. In contrast to the sensor employing the equivalent DNA aptamer, the 2'O-methyl-RNA aptamer sensor lost very little signal and had improved signal-to-noise. We further characterized the EAB sensor drift using unstructured DNA or 2'O-methyl-RNA oligonucleotides. While the two devices drift similarly in vitro in whole blood, the in vivo drift of the 2'O-methyl-RNA-employing device is less compared to the DNA-employing device. Studies of the electron transfer kinetics suggested that the greater drift of the latter sensor arises due to enzymatic DNA degradation. These findings, coupled with advances in the selection of aptamers employing XNA, suggest a means of improving EAB sensor stability when they are used to perform molecular monitoring in the living body.
Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Técnicas Eletroquímicas , Aptâmeros de Nucleotídeos/química , Técnicas Eletroquímicas/métodos , Técnicas Biossensoriais/métodos , Animais , Ratos , Tobramicina/análiseRESUMO
Objetivos: Avaliar e comparar a sensibilidade "in vitro" de bactérias isoladas do olho humano aos aminoglicosídeos gentamicina e tobramicina, e analisar a mudança de sensibilidade ocorrida após quartoze anos de uso.Métodos: Os resultados dos antibiogramas realizados no período de três anos com 887 bactérias (Estudo "A") foram comparados com os resultados obtidos há 14 anos com 124 microrganismos(Estudo"B"), quando tobramicina foi pela primeira vez testada no Brasil para cepas isoladas do olho.Resultados: A eficácia "in vitro" de ambos os antibióticos em relaçäo ao total dos isolamentos positivos foi significantemente mais alta para a tobramicina em ambos os estudos.A comparaçäo dos resultados atuais de sensibilidade com aqueles obtidos há 14 anos demonstra uma menor sensibilidade à tobramicina com relaçäo ao total de bactérias analisadas.Observou-se também um aumento de cepas de Staphylococcus aureus resistentes à tobramicina e apenas uma tendência à diminuiçäo do número de cepas deste microrganismo resistentes à gentamicina.Com relaçäo à Pseudomonas sp, existe uma forte tendência ao aumento de sensibilidade à gentamicina e o aparecimento de cepas resistentes à tobramicina.Discussäo: A introduçäo da tobramicina no tratamento de infecçöes oculares e uma provável diminuiçäo do número de prescriçöes de gentamicina durante estes 14 anos podem ser a causa das modificaçöes da sensibilidade observada do total de isolamentos positivos, principalmente de Staphylococcus aureus e Pseudomonas sp.Para os microrganismos isolados da conjuntiva, o espectro de açäo dos dois antibióticos equivalente.