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Constant temperature reaction-based method for detecting microRNA in solution to be detected

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

Active Publication Date: 2015-10-28
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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  • Constant temperature reaction-based method for detecting microRNA in solution to be detected
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  • Constant temperature reaction-based method for detecting microRNA in solution to be detected

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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

The invention relates to a constant temperature reaction-based method for detecting microRNA in a solution to be detected. The method comprises that an electrode is immersed in a first DNA fragment molecular solution, undergoes a reaction at a temperature of 37 DEG C for 6h, then is immersed in a mercapto hexanol solution, undergoes a reaction for 1h, then is immersed in a silver nanoparticle solution, undergoes a reaction for 1h, then is immersed in a second DNA fragment molecular solution and undergoes a reaction for 0.5h, a microRNA standard liquid is prepared, the electrode is immersed in the microRNA standard liquid, a third DNA fragment molecule, a Klenow fragment and dNTPs are added into the microRNA standard liquid, the electrode undergoes a reaction in the mixed liquid at a temperature of 37 DEG C for 1h, then is immersed in a Nt. BbvCI solution and undergoes a reaction for 1h, wherein in the detection, a linear scanning voltammetry method is used, a standard curve graph is drawn, an electrical signal of the liquid to be detected is acquired by the above processes, and the electrical signal and the standard curve graph are compared so that a microRNA concentration of the liquid to be detected is obtained. The constant temperature reaction-based method has the advantages of simple processes, high sensitivity, good 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 Applications(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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