研究目的
To develop a highly sensitive and selective biosensor for the detection of microRNA-155 using a tripod structure fabricated by triplex DNA and electrogenerated chemiluminescence (ECL) as the detection method.
研究成果
The developed biosensor demonstrated a very low limit of detection (0.10 fM) and a wide linear dynamic range (0.50 fM to 100 pM) for miRNA-155, with excellent specificity, stability, and reproducibility. It was successfully applied to detect miRNA-155 in colon cancer cells, indicating its potential for early disease diagnosis.
研究不足
The study does not mention the biosensor's performance in complex biological matrices other than the colon cancer cell line DLD1. The potential for non-specific binding or interference from other biomolecules in more complex samples is not addressed.
1:Experimental Design and Method Selection:
The biosensor was designed using a tripod structure fabricated by triplex DNA (tsDNA) assembled on electrodeposited reduced graphene oxide (ERGO) as the capture probe. A hybridization chain reaction (HCR) was used for signal amplification by promoting the intercalation of the ECL emitter [Ru(bpy)2(dppz)]2+.
2:2+. Sample Selection and Data Sources:
2. Sample Selection and Data Sources: The target analyte was miRNA-155 from both a standard and a real biological sample (colon cancer cell line DLD1).
3:1). List of Experimental Equipment and Materials:
3. List of Experimental Equipment and Materials: Glassy carbon electrode (GCE), reduced graphene oxide (GO), tri-n-propyl amine (TPrA), [Ru(bpy)2(dppz)]2+, and specific DNA sequences for tsDNA and HCR.
4:Experimental Procedures and Operational Workflow:
The ERGO was electrodeposited on GCE, followed by tsDNA assembly. The biosensor was then treated with target miRNA, H1-DNA and H2-DNA for HCR, and Ru-dppz for ECL signal generation.
5:Data Analysis Methods:
The biosensor's performance was characterized using atomic force microscopy, gel electrophoresis, cyclic voltammetry, and electrochemical impedance spectroscopy. ECL signals were recorded under anodic potential scanning.
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