Detection method for microrna in test solution based on constant temperature reaction

A detection method, a technology of the liquid to be tested, applied in the direction of material electrochemical variables, etc., to achieve the effect of wide application range, low background signal, and simple operation

Active Publication Date: 2018-01-05
SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, it is still a challenge to develop simple and direct analytical techniques with high sensitivity, high specificity, and accurate detection of microRNAs.

Method used

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  • Detection method for microrna in test solution based on constant temperature reaction
  • Detection method for microrna in test solution based on constant temperature reaction
  • Detection method for microrna in test solution based on constant temperature reaction

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Embodiment Construction

[0022] The present invention will be further described in detail below in conjunction with the accompanying drawings, so that those skilled in the art can implement it with reference to the description.

[0023] The detection method of microRNA in the liquid to be tested based on the constant temperature reaction mentioned in this case includes the following steps:

[0024] Step 1) immerse the electrode in a solution containing 0.1 μM of the first DNA fragment molecule, react at 37°C for 6 hours, then immerse the electrode in a solution containing 0.1M mercaptohexanol, and react at 37°C for 1 hour; Wherein, the first DNA fragment molecule is a DNA signal probe, which needs to contain at least one sulfhydryl group and one amino group; It cannot completely cover the entire electrode surface, so it is necessary to put the electrode into the mercaptohexanol solution again, so that the mercapto groups in the mercaptohexanol can fill the position on the electrode surface without DNA...

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Abstract

This case involves a detection method for microRNA in the liquid to be tested based on a constant temperature reaction: immerse the electrode in the first DNA fragment molecule solution, react at 37°C for 6 hours, then immerse in the mercaptohexanol solution for 1 hour; then immerse in the silver nanoparticle solution for reaction 1h, then immerse in the solution of the second DNA fragment molecule and react for 0.5h; prepare microRNA standard solution; then immerse in microRNA standard solution, add the third DNA fragment molecule, Klenow fragment and dNTPs, and react at 37°C for 1h; then immerse in Nt. React in BbvCI solution for 1 h; use linear sweep voltammetry to detect and draw a standard curve; obtain the electrical signal of the test solution according to the above method, and compare it with the standard curve to obtain the concentration of microRNA in the test solution. This case has the advantages of simple operation, high sensitivity, strong specificity, low background signal, and wide application range.

Description

technical field [0001] The invention relates to the field of microRNA detection, in particular to a detection method for microRNA in a test solution based on a constant temperature reaction. Background technique [0002] MicroRNA is a kind of endogenous non-coding RNA with gene regulation function, and its size is about 18-25 nucleotides. microRNA can participate in a variety of regulatory pathways in cells, including development, cell proliferation and apoptosis, organ formation, hematopoietic process, virus defense, metabolism, etc. More and more studies have found that microRNA plays a vital role in the development of tumors and has become an ideal tumor marker. Therefore, quantitative detection of microRNA is of great significance for understanding its biological function, early diagnosis of cancer, development of new drugs and other scientific research projects involving biology. Compared with traditional nucleic acid detection, microRNA has some unique characteristic...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/48
Inventor 缪鹏王弼陡陈锡峰孙海旋唐玉国
Owner SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI
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