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An Electrogenerated Chemiluminescence Biosensor using a tripod probe for the Highly Sensitive Detection of MicroRNA

DOI:10.1021/acs.analchem.8b04271 期刊:Analytical Chemistry 出版年份:2018 更新时间:2025-09-04 15:30:14
摘要: A novel probe for highly sensitive detection of microRNA, that enhanced the helix accessibility and yielded good assembling without backfilling, was developed using a tripod structure fabricated by triplex DNA. A layer of triplex DNA assembled on electrodeposited reduced graphene oxide was used as the capture probe and a subsequent hybridization chain reaction that promoted the efficient intercalation of the electrogenerated chemiluminescence(ECL) emitter [Ru(bpy)2(dppz)]2+ (bpy = 2,2′-bipyridine, dppz = dipyrido[3,2-a: 2′,3′-c]phenazine) was used an analytical signal amplifier. The fabricated biosensor was examined with an anodic ECL mode using tri-n-propyl amine as the coreactant. The construction of the biosensor was systematically characterized with various techniques including atomic force microscopy, gel electrophoresis, cyclic voltammetry, and electrochemical impedance spectroscopy, and its performance was optimized under a variety of experimental conditions especially the concentration of each reagent as well as its incubation time. Under the optimal experimental conditions, the reported biosensor showed a very low limit of detection of 0.10 fM (S/N = 3) and a wide linear dynamic range covering from 0.50 fM to 100 pM towards microRNA-155, with excellent specificity, stability, and reproducibility. Finally, the biosensor was successfully applied to detect the amount of microRNA-155 extracted from the colon cancer cell line DLD1, demonstrating its potential applications in sensitive detection of biological samples in early diagnosis of diseases.
作者: Liping Lu,Jiaxing Wang,Wujian Miao,Xiayan Wang,Guangsheng Guo
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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.

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.

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.

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