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Multiple microRNA detection method of silver nano-cluster based on entropy-driven amplification system as template

A silver nanocluster and amplification system technology, applied in the field of life analysis chemistry, can solve problems that need to be further developed, achieve excellent biocompatibility, simple preparation method, and avoid cross-band interference

Pending Publication Date: 2020-12-29
TIANJIN UNIV OF TRADITIONAL CHINESE MEDICINE
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the multiple detection of DNA and adenosine phosphate has been realized as a signal output method, but the multiple detection of indicator miRNA needs to be further developed
Therefore, label-free, spectrally flexible and tunable optical labels as signal transduction methods are another challenge for multiplex miRNA detection.

Method used

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  • Multiple microRNA detection method of silver nano-cluster based on entropy-driven amplification system as template
  • Multiple microRNA detection method of silver nano-cluster based on entropy-driven amplification system as template
  • Multiple microRNA detection method of silver nano-cluster based on entropy-driven amplification system as template

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Experimental program
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Effect test

Embodiment 1

[0027] Design of miRNA detection system sequence:

[0028] The DNA and microRNA primers used in the examples were all synthesized by Sangon Bioengineering (Shanghai) Co., Ltd. and purified by high performance liquid chromatography. The primer sequences are shown in the table below:

[0029]

[0030]

Embodiment 2

[0032] Preparation of triplex complexes SY and SR in an entropy-driven catalytic amplification system:

[0033] Mix primers SY-1, SY-2 and SY-3 at equal concentrations (10 μM) in 20 mM phosphate buffer solution (including 10 mM magnesium acetate), heat at 95°C for 15 minutes, and slowly cool down to room temperature to obtain the triplex complex SY .

[0034] Mix primers SR-1, SR-2 and SR-3 at equal concentrations (10 μM) in 20 mM phosphate buffer solution (including 10 mM magnesium acetate), heat at 95°C for 15 minutes, and slowly cool down to room temperature to obtain the triple-stranded complex SR .

Embodiment 3

[0036] Preparation of silver nanoclusters with three-chain complexes SY and SR as templates:

[0037] Take 80 μL of the prepared three-chain complex SY and SR solutions and dilute them with phosphate buffer (final reaction concentration is 2 μM), add 4.8 μL of 1 mM silver nitrate solution, mix well, and place in the dark at room temperature for 15 minutes. Continue to add 4.8 μL of 1mM sodium borohydride solution to the reaction solution, mix well, and react in the dark at room temperature for 6 hours, and finally obtain yellow-emitting silver nanocluster SY-Ag NCs and red-emitting silver nanocluster SR-Ag NCs.

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Abstract

The present invention provides a multiple microRNA detection method of a silver nano-cluster based on an entropy-driven amplification system as a template and belongs to the technical field of life analysis chemistry. An entropy-driven catalytic reaction is used as an amplification element, the silver nano-cluster prepared by using a three-chain compound in the catalytic reaction as a template isused as an optical label, and the multiple microRNA detection is simply, quickly and sensitively carried out. microRNA triggers branch migration and chain substitution of the three-chain compound forming the luminescent silver nano-cluster, so that a structure of the silver nano-cluster is changed, fluorescence is sharply reduced, a reduction degree is positively correlated with concentration of the microRNA, and meanwhile, the microRNA is released for a new round of detection to generate changes of amplified fluorescence signals. The detection method economically and efficiently realizes independent detection and simultaneous multiple detection of miRNA-141 and miRNA-155.

Description

technical field [0001] The invention belongs to the technical field of life analysis chemistry, and in particular relates to a multiple microRNA detection method based on an entropy-driven amplification system using silver nanoclusters as templates. Background technique [0002] MicroRNA (miRNA for short) is a short, non-coding RNA with a length of about 18-25 nucleotides. miRNAs play important roles in regulating various developmental, pathological and physiological processes. Abnormal expression of miRNAs is closely related to many human diseases, including cancer, neurological diseases, cardiovascular diseases, kidney and liver diseases, etc. In recent years, miRNA has become a very valuable biomarker, which is of great significance in the early diagnosis, prognosis and treatment of diseases. In addition, the occurrence and progression of diseases are often closely related to multiple miRNAs, and the detection of a single miRNA may lead to wrong conclusions and even mis...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/682C12Q1/6806
CPCC12Q1/682C12Q1/6806C12Q2525/207C12Q2563/107C12Q2537/143C12Q2537/119C12Q2563/137C12Q2563/155
Inventor 李凤云龚珈苧邱峰
Owner TIANJIN UNIV OF TRADITIONAL CHINESE MEDICINE
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