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1.
Biosens Bioelectron ; 194: 113618, 2021 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-34530373

RESUMO

Carcinoembryonic antigen (CEA) is an important malign tumor marker. In this study, a simple, label-free and antibody-free aptasensor was fabricated based on a multifunctional dendrimer-like DNA nanoassembly. The DNA nanoassembly was embedded with multiple G-quadruplex DNAzyme motifs and a hanging CEA aptamer motif. It was prepared from short DNA sequences by autonomous-assembly. The aptasensor was prepared simply by self-assembly of a capture DNA (cpDNA) on a gold electrode, followed by hybridization with a CEA aptamer (AptGAC-P). CEA as a model target was detected through competitive binding of CEA with AptGAC-P, exposing cpDNA to bind with the DNA nanoassembly. The detection process only contains 2 incubation steps. The high load of G-quadruplex DNAzyme motifs and their catalytic activity resulted in an amplified and label-free differential pulse voltammetry (DPV) electrochemical signal. The peak current correlated linearly with the CEA concentration, with a linear range of 2-45 ng mL-1, and an LOD value of 0.24 ng mL-1. The aptasensor showed high specificity and reproducibility, and retained 96.5% of detection signal intensities after 31 days of storage. The recovery rates for spiked CEA in human serum were within 100 ± 5%, and the coincidence rates for clinical human serum samples with ELISA kits were 80.7-111%. Conceivably, possessing simplicity, sensitivity, reproducibility, storage stability, and accuracy, the aptasensor should be a very prominent and applicable tool for clinical CEA detection and cancer diagnosis, and is promisingly applicable as a platform for detecting other targets of interests.


Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , DNA Catalítico , Dendrímeros , Antígeno Carcinoembrionário , Catálise , DNA , Humanos , Reprodutibilidade dos Testes
2.
Biosens Bioelectron ; 141: 111452, 2019 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-31252259

RESUMO

A stable and sensitive electrochemical acetylcholinesterase (AChE) biosensor for detection of organophosphorus pesticides (OPs) was developed by doping Au nanorods (AuNRs)@mesoporous SiO2 (MS) core-shell nanoparticles into CS/TiO2-CS (CS denotes for chitosan) immobilization matrix. AuNRs@MS core-shell nanoparticles were synthesized and characterized. The doping and the biosensor fabrication process were probed and confirmed by scanning electron microscopy and electrochemistry techniques. The doping conditions were optimized. The matrix both before and after AChE immobilization had a mesoporous nanostructure. The nanoparticles dispersed homogeneously within the matrix. The doping significantly enhanced the electro-conductivity of the TiO2-CS hydrogel, and dramatically improved the bioelectrocatalytic activity and OPs detection sensitivity of the AChE immobilized matrix. The detection linear ranges for both dichlovos (DDVP) and fenthion were from 0.018 µM (4.0 ppb) to 13.6 µM, and the limit of detection (LOD) was 5.3 nM (1.2 ppb) and 1.3 nM (0.36 ppb), respectively. The biosensor exhibited high reproducibility and accuracy in detecting OPs spiked vegetable juice samples. In addition, it exhibited very high detection stability and storage stability. The developed AChE biosensor was provided to be a promisingly applicable tool for OPs detection with high reliability, simplicity, and rapidness.


Assuntos
Técnicas Biossensoriais/métodos , Ouro/química , Hidrogéis/química , Nanotubos/química , Compostos Organofosforados/análise , Praguicidas/análise , Acetilcolinesterase/química , Animais , Quitosana/química , Electrophorus , Enzimas Imobilizadas/química , Proteínas de Peixes/química , Limite de Detecção , Nanopartículas/química , Dióxido de Silício/química , Titânio/química
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