Double-signal miRNA-21 detection method based on three-dimensional DNA Walker and MOF-Fe (II) induced generation of Turnskill blue

A miRNA-21 and detection method technology, which is applied in the direction of measuring devices, instruments, and material analysis through electromagnetic means, to achieve the effects of improving detection sensitivity, high reactivity, and wide linear range

Active Publication Date: 2021-11-09
JIANGXI NORMAL UNIV
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Problems solved by technology

Compared with other methods, the electrochemical and temperature-based sensing platform has obvious advantages such as simplicity, low cost and high sensitivity, which can realize the sensitive detection of miRNA, but the existing technology uses three-dimensional DNA Walker and MOF-Fe( II) Inducing Teng's blue production to realize miRNA-21 electrochemical and photothermal detection methods have not been reported yet

Method used

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  • Double-signal miRNA-21 detection method based on three-dimensional DNA Walker and MOF-Fe (II) induced generation of Turnskill blue
  • Double-signal miRNA-21 detection method based on three-dimensional DNA Walker and MOF-Fe (II) induced generation of Turnskill blue
  • Double-signal miRNA-21 detection method based on three-dimensional DNA Walker and MOF-Fe (II) induced generation of Turnskill blue

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Embodiment 1

[0056] (1) First prepare 10mL of 0.57M FeAc 2 aqueous solution; subsequently, accurately weigh 0.9 g of BDC-NH 2 , added to 30mL of DMF, and ultrasonically treated for about 10min to fully dissolve it; while stirring, FeAc 2 Aqueous solution was added to BDC-NH 2 DMF solution, stirred for 15 min, transferred to an autoclave, and allowed to react at 50°C for 1 hour; after cooling to ambient temperature, the product was collected by centrifugation, and washed three times with DMF and absolute ethanol; finally, the The red-brown product obtained was dried overnight in a vacuum oven.

[0057] (2) Ultrasonically disperse 1 mg of MOF-Fe(II) powder prepared in step (1) in 2 mL of methanol, and drop 4.5 L onto the surface of GCE. After drying at room temperature, the MOF-Fe(II) / GCE electrode was obtained.

[0058] (3) Add 10 μL of avidin-modified MB (Strep-MB, 5 mg mL -1 ) with 10 μL biotin-modified hairpin H 1 (2μM) mixed and incubated at 37°C for 30min. For magnetic separatio...

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Abstract

The invention discloses a double-signal miRNA-21 detection method based on three-dimensional DNA Walker and MOF-Fe (II) induced generation of Turnskill blue. The method comprises the following steps: mixing and incubating avidin modified MB and a biotin modified hairpin strand H1, and then mixing and incubating with a hairpin strand H2-Au NP-GOx probe and a miRNA-21 solution, so as to obtain a DNA Walker product; adding the DNA Walker product into a glucose solution to carry out catalytic reaction, and carrying out magnetic separation to obtain a solution containing hydrogen peroxide; and enabling the MOF-Fe (II) to react with a solution containing hydrogen peroxide and induce generation of TB on an electrode to realize electrochemical detection, or enabling the MOF-Fe (II) solution containing hydrogen peroxide to induce generation of TB in the solution to realize photo-thermal detection, so that the method can be used for dual-signal detection of miRNA, and is low in miRNA target object detection lower limit and high in selectivity.

Description

technical field [0001] The invention relates to a detection method of miRNA-21, specifically a dual-signal miRNA-21 detection method based on three-dimensional DNA Walker and MOF-Fe(II)-induced instant generation of Teng's blue, which belongs to electrochemical and photothermal biological analysis field. Background technique [0002] MicroRNA (miRNA) is a class of short non-coding sequences, 18-25 nucleotides in length, which play a vital role in various biological processes, such as: gene expression, transcription, cell proliferation, differentiation, Apoptosis and hematopoiesis. In addition, abnormal expression of miRNAs will lead to cancer formation, invasion and metastasis. In recent years, many reported analytical methods combined with biological analysis have been successfully used for rapid quantitative or semi-quantitative analysis of miRNA, including electrochemical, fluorescence, colorimetric, surface plasmon resonance, and electrochemiluminescence-based biologic...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/30G01N27/327G01N27/48
CPCG01N27/308G01N27/3277G01N27/3276G01N27/3278G01N27/305G01N27/48Y02A50/30
Inventor 汤娟程宏丽刘丽萍高珊
Owner JIANGXI NORMAL UNIV
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