【摘 要】
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Exosomal miRNAs,as potential biomarkers in liquid biopsy for cancer early diagnosis,have aroused widespread concern.Herein,an electrochemical biosensor based on DNA “nano-bridge” was designed and applied to detect exosomal microRNA-21 (miR-21) derived fro
【机 构】
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College of Life Sciences,Fujian Agriculture and Forestry University,Fuzhou 350002,China;Department o
论文部分内容阅读
Exosomal miRNAs,as potential biomarkers in liquid biopsy for cancer early diagnosis,have aroused widespread concern.Herein,an electrochemical biosensor based on DNA “nano-bridge” was designed and applied to detect exosomal microRNA-21 (miR-21) derived from breast cancer cells.In brief,the target miR-21 can specifically open the hairpin probe 1(HP1) labeled on the gold electrode (GE) surface through strand displacement reaction.Thus the exposed loop region of HP1 can act as an initiator sequence to ac-tivate the hybridization chain reaction (HCR) between two kinetically trapped hairpin probes:HP2 immo-bilized on the GE surface and biotin labeled HP3 in solution.Cascade HCR leads to the formation of DNA“nano-bridge” tethered to the GE surface with a great deal of “piers”.Upon addition of avidin-modified horseradish peroxidase (HRP),numerous HRP were bound to the formed “nano-bridge” through biotin-avidin interaction to arouse tremendous current signal.In theory,only a single miR-21 is able to trigger the continuous HCR between HP2 and HP3 until all of the HP2 are exhausted.Therefore the proposed biosensor achieved ultrahigh sensitivity toward miR-21 with the detection limit down to 168 amol/L,as well as little cross-hybridization even at the single-base-mismatched level.Successful attempts were also made in the detection of exosomal miR-21 obtained from the MCF-7 of breast cancer cell line.To our knowledge,this is the first attempt to built horizontal DNA nano-structure on the electrode surface for exosomal miRNAs detection.In a word,the high sensitivity,selectivity,low cost make the proposed method hold great potential application for early point-of-care (POC) diagnostics of cancer.
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