We proposed a simple and sensitive strategy for the detection of microRNAs (miRNAs) by converting homogeneous assay into surface-tethered electrochemical analysis. Specifically, the biotinylated detection probes (biotin-DNA-biotin) can trigger the in-situ assembly of tetrameric streptavidin (SA) proteins on an electrode surface via the SA-biotin interactions. The (SA-biotin-DNA-biotin) assemblies electrically insulated the electrode interface, thereby blocking the electron transfer of [Fe(CN)]. When the probe was hybridized with the target miRNA, it would be cleaved into small fragments (denoted as biotin-DNA) by duplex-specific nuclease (DSN). The released target miRNA can enter into the next hybridization-enzymolysis cycle, thus leading to the generation of considerable amounts of biotin-DNA fragments. The released biotin-DNA competed with the detection probe to bind SA, thus limiting the in-situ formation of (SA-biotin-DNA-biotin) assemblies. The surface-tethered electrochemical analysis by the dual signal amplification of DSN and (SA-biotin-DNA-biotin) assemblies has been used for the determination of miRNAs in cell lysate with a satisfactory result. The method showed a detection limit down to 10 aM. The "one-step" immobilization-free strategy can be used to design novel biosensors for the detection of other biomarkers.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1016/j.aca.2021.338199 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Electrochemical detection of poly(ADP-ribose) polymerase-1 with silver nanoparticles as signal labels by integrating the advantages of homogeneous reaction with surface-tethered detection.
Talanta
January 2025
Henan Province Key Laboratory of New Opto-electronic Functional Materials, College of Chemistry and Chemical Engineering, Anyang Normal University, Anyang, Henan, 455000, China. Electronic address:
Poly(ADP-ribose)polymerase-1 (PARP1) could be activated by binding to nucleic acids with specific sequences, thus catalyzing the poly-ADP-ribosylation (PARylation) of target proteins including PARP1 itself. Most of the previously reported electrochemical methods for the determination of PARP1 were relied on the electrostatic interactions, which required the pre-immobilization of DNA on an electrode for the capture of PARP1. Herein, we reported an "immobilization-free" electrochemical strategy for the assays of PARP1 on the basic of avidin-biotin interaction.
View Article and Find Full Text PDFSemiautomated Electrochemical Melting Curve Analysis Device for the Detection of an Osteoporosis Associated Single Nucleotide Polymorphism in Blood.
Anal Chem
September 2023
INTERFIBIO Research Group, Departament d'Enginyeria Química, Universitat Rovira i Virgili, 43007 Tarragona, Spain.
Article Synopsis
- The detection of single nucleotide polymorphisms (SNPs) is crucial for clinical diagnostics, pharmacogenomics, and forensics, particularly for identifying genetic risks like those associated with osteoporosis.
- A semiautomated system using solid-phase electrochemical melting curve analysis (éMCA) was developed to identify alleles at specific SNP sites related to bone fracture risks by employing asymmetric isothermal recombinase polymerase amplification.
- The proof-of-concept utilized a microfluidic device with a multielectrode array, allowing for the effective detection of SNP hetero/homozygosity, particularly at the osteoporosis-related SNP site rs2741856 in real patient samples.
Probing charge transfer through antifouling polymer brushes by electrochemical methods: The impact of supporting self-assembled monolayer chain length.
Anal Chim Acta
October 2023
J. Heyrovský Institute of Physical Chemistry of the Czech Academy of Sciences, Dolejškova 3, 182 23, Prague, Czech Republic. Electronic address:
Ultrathin surface-tethered polymer brushes represent attractive platforms for a wide range of sensing applications in strategically vital areas such as medicine, forensics, or security. The recent trends in such developments towards "real world conditions" highlighted the role of zwitterionic poly(carboxybetaine) (pCB) brushes which provide excellent antifouling properties combined with bio-functionalization capacity. Highly dense pCB brushes are usually prepared by the "grafting from" polymerization triggered by initiators on self-assembled monolayers (SAMs).
View Article and Find Full Text PDFPractical tips and new trends in electrochemical biosensing of cancer-related extracellular vesicles.
Anal Bioanal Chem
March 2023
Department of Chemistry Ugo Schiff, University of Florence, Via Della Lastruccia 3, 50019, Sesto, Fiorentino, Italy.
To tackle cancer and provide prompt diagnoses and prognoses, the constantly evolving biosensing field is continuously on the lookout for novel markers that can be non-invasively analysed. Extracellular vesicles (EVs) may represent a promising biomarker that also works as a source of biomarkers. The augmented cellular activity of cancerous cells leads to the production of higher numbers of EVs, which can give direct information on the disease due to the presence of general and cancer-specific surface-tethered molecules.
View Article and Find Full Text PDFSurface-tethered electrochemical biosensor for telomerase detection by integration of homogeneous extension and hybridization reactions.
Talanta
February 2023
College of Chemistry and Chemical Engineering, Anyang Normal University, Anyang, Henan 455000, People's Republic of China. Electronic address:
The general electrochemical biosensors for telomerase detection require the immobilization of primers on the electrode surface for telomeric extension and hybridization reactions. However, immobilization of primers may suffer from the challenges of hindrance effect and configuration freedom, thus reducing the extension and hybridization efficiency. Herein, we developed a sensitive electrochemical biosensor for telomerase detection by integration of homogeneous extension and hybridization reactions and surface-tethered detection.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!