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Micro RNA detection method based on one-step triggered branched DNA nanostructure

A nanostructure and tiny technology, applied in the direction of biochemical equipment and methods, microbial measurement/testing, etc.

Active Publication Date: 2020-08-18
CHONGQING TECH & BUSINESS UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there is no precedent for the detection of microRNA by hybridization chain reaction (HCR).

Method used

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  • Micro RNA detection method based on one-step triggered branched DNA nanostructure
  • Micro RNA detection method based on one-step triggered branched DNA nanostructure
  • Micro RNA detection method based on one-step triggered branched DNA nanostructure

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Embodiment Construction

[0028] The present invention will be described in further detail below in conjunction with the embodiments.

[0029] Such as figure 1 As shown, the principle of the present invention is: H1 is fixed on the surface of the gold electrode through the mercapto groups on the five initiator groups. MCH is then used to block the surface and reduce nonspecific adsorption, thereby increasing hybridization efficiency and accessibility of surface-bound hairpin DNA. Target microRNAs can hybridize to H1, thereby exposing the ends of H1. The end of the H1 is complementary to the toe and stem region of the H2, resulting in an opening of the H2 and exposing the end of the H2. The end of H2 can unfold H3 and release the end of H3. The release end of H3 is in the same order as the active part of H1. Thus, HCR-1 between H2 and H3 can be triggered to form a long nick of dsDNA flanked by some capture probes. Then, capture probes flanking the duplex can prime HCR-2 between two alternating hair...

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Abstract

The invention provides a target micro RNA induced-based branched DNA nanostructure and develops a simple and ultra-sensitive micro RNA detection method based on a target micro RNA induced branched DNAnanostructure. Short DNA single-stranded tails on the two sides of the double-stranded body can use hairpin DNA4 and hairpin DNA5 to start second-stage HCR amplification; a plurality of long double-stranded bodies are generated on the two sides of the formed long double-helix DNA, so that the formed branched chain DNA nano structure can carry a large amount of methyl blue, an electrochemical signal can be effectively enhanced, and a low detection limit is realized. More importantly, the strategy provides simple operation, good reliability and ultrahigh sensitivity. The method has been successfully applied to human serum samples, and shows satisfactory results and hopeful future. Importantly, the one-stop method can greatly reduce operation steps and avoid system errors caused by a plurality of operation steps.

Description

technical field [0001] The invention relates to the detection field of microRNA, in particular to the field of microRNA detection method based on one-step triggered branched DNA nanostructure. Background technique [0002] MicroRNA is a class of small non-coding RNA with a length of about 19-23nt. It has emerged as a post-transcriptional regulator that plays an important role in every fundamental biological process such as proliferation, differentiation and apoptosis. Many human microRNAs have been reported to be expressed throughout the genome in disease etiology including, for example, cancer, cardiovascular disease, and Alzheimer's disease. Studying the expression of microRNA is of great value for the diagnosis and prognosis of early cancer. Therefore, it is highly desirable to develop a method for microRNA detection, especially for complex clinical samples and single cells. Several existing technologies have shown potential for microRNA clinical diagnosis, including N...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/682C12Q1/6825
CPCC12Q1/682C12Q1/6825C12Q2525/301C12Q2565/607
Inventor 云雯熊政委哈霞吴虹李宁
Owner CHONGQING TECH & BUSINESS UNIV
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