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Nucleotide sequence for detecting mircoRNA content in samples and detecting method

A technology in nucleotide sequence and samples, which is applied in the field of detecting microRNAs in samples using gold and silver nanosphere technology, can solve the problems of sensitivity and specificity to be improved, and achieve the effect of improving sensitivity, high sensitivity, and high sensitivity

Active Publication Date: 2009-09-23
HARBIN MEDICAL UNIVERSITY
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Problems solved by technology

However, its sensitivity and specificity still need to be improved

Method used

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  • Nucleotide sequence for detecting mircoRNA content in samples and detecting method
  • Nucleotide sequence for detecting mircoRNA content in samples and detecting method
  • Nucleotide sequence for detecting mircoRNA content in samples and detecting method

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preparation example Construction

[0053] 1. Sample preparation and total RNA extraction were performed using conventional methods and reagents;

[0054] 2. Hybridization: mix the sample, capture probe, signal probe and nano-gold labeling probe, transfer the mixture system to each streptavidin-coated microplate hole and incubate, the nano-gold labeling probe The needle is obtained by mixing a universal T probe with a nano-gold solution;

[0055] 3. Quantitative detection of microRNA with nano-gold-labeled probes: Add silver enhancement solution to each well of the aforementioned microplate plate, react at room temperature and dark conditions until a gray signal appears, then rinse with ultrapure water to terminate the reaction, and place the plate into a microplate reader In the process, single-wavelength scanning is used to detect the A value of the solution in real time through a microplate reader.

[0056] Wherein, the nanogold can be purchased from commercial sources (such as purchased from Sino-American B...

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Abstract

The invention provides a nucleotide sequence and a kit for detecting mircoRNA content in samples as well as a mircoRNA detecting method which is based on the complementary combination of nucleic acid base pairing identification and utilizes silver-staining enhanced nano gold to mark a probe, the method is simple and practicable and can obtain detection signals with high sensitivity and high specificity without expensive equipment of radioactive labels, fluorescence and the like.

Description

technical field [0001] The invention relates to a nucleotide sequence and a detection method for detecting the content of microRNAs in samples, in particular to a method for detecting microRNAs in samples using gold and silver nanosphere technology. Background technique [0002] Cardiovascular disease has become the number one killer of human health and life. Acute myocardial infarction is a common emergency and critical illness in the emergency department, but the treatment effect for this disease is not satisfactory. One of the important reasons is that the prognosis of myocardial infarction and the occurrence of complications cannot be correctly judged in time, so that appropriate measures cannot be taken. Timely treatment plans and measures. Malignant arrhythmia is one of the serious complications of acute myocardial infarction. It is a difficult point in clinical treatment and a hot spot in this research field. It has serious consequences and high mortality. Early dete...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11G01N33/53
Inventor 杨宝峰王志国吕延杰张荣初文峰
Owner HARBIN MEDICAL UNIVERSITY
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