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1.
Anal Bioanal Chem ; 414(22): 6441-6453, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-35788872

RESUMO

Hepatocellular carcinoma (HCC) is the second leading cause of cancer-related deaths in China. Glypican-3 (GPC3) is a specific antigen related to HCC, which is widely used in clinical detection as a reliable marker of HCC. In this paper, a highly sensitive homogeneous apatasensor was designed for GPC3 detection based on fluorescence resonance energy transfer (FRET) where the GPC3 aptamer labelled gold carbon dots (AuCDs-GPC3Apt) are used as a donor and magnetic graphene oxide (Fe3O4/GO) nanosheets are used as an acceptor. A one-step hydrothermal method was used to synthesize AuCDs to provide sufficient fluorescence. The FRET phenomenon exists between AuCDs-GPC3Apt and Fe3O4/GO, which weakens the fluorescence intensity of the whole system. When the target GPC3 is added to the FRET system, the fluorescent AuCDs-GPC3Apt binds to the GPC3 and forms a folded structure, which leads to AuCDs-GPC3Apt separation from Fe3O4/GO nanosheets. The Fe3O4/GO is then magnetically separated so that the fluorescence of free labelled AuCDs-GPC3Apt is restored. Under the optimum conditions, the fluorescence recovery rate is linearly correlated with the concentration of GPC3 (5-100 ng·mL-1) and the detection limit is 3.01 ng·mL-1 (S/N = 3). This strategy shows recoveries from 98.76 to 101.29% in real human serum samples and provides an immediate and effective detection method for the quantification of GPC3 with great potential applications for early diagnosis of HCC. A sensitive homogeneous FRET-based apatasensor was designed for GPC3 detection where the AuCDs-GPC3Apt is a donor and Fe3O4/GO nanosheets are an acceptor. The GPC3 fluorescent aptasensor combines wider output range with low cost, high specificity, and good anti-interference.


Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Carcinoma Hepatocelular , Grafite , Neoplasias Hepáticas , Aptâmeros de Nucleotídeos/química , Técnicas Biossensoriais/métodos , Carbono/química , Carcinoma Hepatocelular/diagnóstico , Detecção Precoce de Câncer , Transferência Ressonante de Energia de Fluorescência/métodos , Glipicanas , Ouro/química , Grafite/química , Humanos , Limite de Detecção , Neoplasias Hepáticas/diagnóstico
2.
Bioelectrochemistry ; 147: 108204, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-35839688

RESUMO

Diabetes is one of metabolic diseases affecting major human health. The early diagnosis and treatment of diabetes have significant benefits. 1,5-anhydroglucitol (1,5-AG) accurately reflects a patient's average blood glucose level for the past 3-7 days and becomes a promising marker for real-time detection of diabetes. In this study, a novel biosensor for determination 1,5-AG is constructed using reduce graphene oxide-carboxymethylated chitosan-hemin@platinum nanocomposites (rGO-CMC-H@Pt NCs) nanozyme and pyranose oxidase (PROD) enzyme as the electrochemical biosensing platform. The rGO-CMC-H@Pt NCs nanozyme has good electro-conductibility, high specific surface area, and admirable peroxide-like catalysis effect to enhance the electrochemical response. 1,5-AG is catalyzed by PROD and produces hydrogen peroxide (H2O2), which in turn can be decomposed by rGO-CMC-H@Pt NCs and produce a current signal recorded by differential pulse voltammetry (DPV) technique. Under optimal conditions, the response currents have a linear relationship in the 1,5-AG concentration of 0.1-2.0 mg/mL with R2 of 0.9869. The sensitivity is 2.1895 µA/µg·mL-1 and the limit of detection (LOD) is 38.2 µg/mL (S/N = 3). In addition, the specificity, reproducibility, stability and recovery (94.5-107.6%) of 1,5-AG biosensors all exhibit good performance. Therefore, the designed 1,5-AG biosensor has a good effect and can be used for the diagnosis of diabetes.


Assuntos
Técnicas Biossensoriais , Quitosana , Grafite , Técnicas Biossensoriais/métodos , Citocromo P-450 CYP2B1 , Desoxiglucose , Técnicas Eletroquímicas/métodos , Hemina , Humanos , Peróxido de Hidrogênio , Limite de Detecção , Platina , Reprodutibilidade dos Testes
3.
Biochem Biophys Res Commun ; 610: 30-34, 2022 06 25.
Artigo em Inglês | MEDLINE | ID: mdl-35430449

RESUMO

Circular RNAs (circRNAs) are a class of noncoding RNAs generated by a specific type of RNA alternative splicing called backsplicing through various mechanisms. Recently, thousands of circRNAs have been identified by high-throughput RNA sequencing technologies and bioinformatics analysis. However, the functions of the majority have not been fully elucidated yet. Different tools, such as in situ hybridization, can help visualize the spatial temporal distribution of circRNA molecules, thus assisting the understanding of their biological and physiological functions. Here, we present a simple and straightforward method based on padlock probe hybridization and rolling circle amplification (RCA) for in situ detection of circRNAs. We compared our method with the commercially available BaseScope assay for the detection of Cdr1as in the mouse brain tissue. The result showed that the two methods have achieved comparable detection efficiency, thus demonstrating our padlock probe assay as an alternative yet simple circRNA in situ detection method for the research community.


Assuntos
Técnicas de Amplificação de Ácido Nucleico , RNA Circular , Animais , Hibridização In Situ , Camundongos , Técnicas de Amplificação de Ácido Nucleico/métodos
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