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Serological biomarker miR-19a for detecting coronary heart disease and application of serological biomarker miR-19a

A mir-19a, biomarker technology, applied in the field of coronary heart disease biomarkers, can solve the problems of complex conditions, unclear pathogenesis, rare reports on the mechanism of coronary heart disease system, etc., and achieve the effect of accurate prediction

Inactive Publication Date: 2015-01-14
雷桅
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

More and more evidence shows that inflammation plays an important role in the pathogenesis of coronary heart disease, and its pathogenesis has obvious family aggregation. A large number of genetic susceptibility genes play an important role in the occurrence and development of the disease. The involved The conditions are very complicated, and the exact pathogenesis of the disease is still unclear, which limits the discovery of effective susceptibility genes for the diagnosis and treatment of coronary heart disease
miRNA widely mediates the post-transcriptional regulation of genes, and has been proven to play a variety of roles in cardiovascular diseases, which may reflect the susceptibility of the heart and blood vessels to injury, and the dependence on persistent cardiovascular function, etc., and the current main The research focuses on the relationship between miRNA and arrhythmia, cardiac hypertrophy, angiogenesis, and cardiomyocyte apoptosis, and the systemic mechanism of action on coronary heart disease is rarely reported.

Method used

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  • Serological biomarker miR-19a for detecting coronary heart disease and application of serological biomarker miR-19a
  • Serological biomarker miR-19a for detecting coronary heart disease and application of serological biomarker miR-19a
  • Serological biomarker miR-19a for detecting coronary heart disease and application of serological biomarker miR-19a

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Subject group

[0021] The test subjects were patients diagnosed and treated in the hospital. Inclusion criteria for patients with coronary heart disease: previous history of myocardial infarction, or at least one coronary artery stenosis ≥ 50% in coronary angiography; age greater than 18 and less than 80 years old. Exclusion criteria: angina pectoris at rest; patients with severe heart failure (EF < 35%); patients with severe valvular disease or cardiomyopathy. A total of 180 cases in the coronary heart disease group and 169 cases in the non-coronary heart disease group were selected, and all subjects signed informed consent.

[0022] Table 1 Basic information of cases

[0023]

Embodiment 2

[0025] Plasma miRNA extraction

[0026] (1) Collect 1 ml of fresh blood from the patient, place it in an anticoagulant tube, centrifuge at 3000 rpm for 10 minutes at 4°C, and collect the upper plasma;

[0027] (2) Take 200 μl of plasma, add an equal volume of lysate MZ, shake and mix for 30 s;

[0028] (3) Leave at room temperature for 5 minutes to completely separate the protein complex;

[0029] (4) Centrifuge at 12,000 rpm for 10 min at room temperature, take the supernatant, and transfer it to a new RNase-free centrifuge tube;

[0030] (5) Add 200 μl of chloroform, shake vigorously for 15 s, and place at room temperature for 5 min;

[0031] (6) After centrifugation at 12,000 rpm at room temperature for 15 minutes, the sample will be divided into 3 layers: a yellow organic phase, an intermediate layer, and a colorless aqueous phase. RNA exists in the aqueous phase. Transfer the aqueous phase to a new tube;

[0032] (7) Measure the volume of the transfer solution, slow...

Embodiment 3

[0041] cDNA first-strand synthesis of miRNA by reverse transcription

[0042] (1) Add poly(A) to the 3' end of miRNA

[0043] 1) Put the 200 μl reaction tube without RNase on ice and add the following reagents (E.coliPoly(A)

[0044] Polymerase was added last).

[0045]

[0046] 2) Gently aspirate and mix the above reaction solution with a pipette, centrifuge briefly and react at 37°C for 60 min.

[0047]

[0048] (2) Reverse transcription reaction of poly(A) modified miRNA

[0049] 1) Prepare the reaction solution according to the following components.

[0050]

[0051] 2) Gently aspirate and mix the above reaction solution with a pipette, centrifuge briefly and react at 37°C for 60 min. The synthesized cDNA reaction solution was stored at -20°C.

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Abstract

The invention relates to a serological biomarker miR-19a for detecting the coronary heart disease and application of the serological biomarker miR-19a, belonging to the fields of laboratory medicines, clinical medicines, biotechnology, biochemistry and molecular biology. The miRNA sequence is as follows: miR-19a: UGUGCAAAUCUAUGCAAAACUGA. The invention further discloses application of the miR-19a. The miR-19a is used for preparing a reagent product for screening the coronary heart disease or assisting coronary heart disease pathological authentication, biochemical detection and clinical diagnosis. The reagent product comprises a primer system and a primer probe system, wherein the primer system consists of cDNA amplification primers of the miR-19a; the primer probe system consists of the cDNA amplification primers of the miR-19a and a probe. The biomarker miR-19a molecule is closely related with the occurrence and the development of the coronary heart disease; and serving as the biomarker for predicating the occurrence of the coronary heart disease and judging the development degree of the coronary heart disease, compared with the existing normal pathological type diagnosis for an attack of the coronary heart disease and prognosis, the serological biomarker miR-19a can be used for predicating the early-stage morbidity and the malignancy degree of the coronary heart disease individually and accurately.

Description

technical field [0001] The present invention relates to a serological biomarker miR-19a for detecting coronary heart disease and its application, in particular to a biomarker for coronary heart disease—serum miR-19a molecule and its role in coronary heart disease screening and clinical diagnosis The application belongs to the fields of laboratory medicine, clinical medicine, biotechnology, biochemistry and molecular biology. Background technique [0002] Coronary heart disease (CAD) has high morbidity and mortality worldwide, especially in some developing countries. According to the prediction of WHO, until 2030, 23,600,000 people will die of cardiovascular diseases every year. The pathological process of coronary heart disease is complex, and it is generally considered to be the result of the joint action of genetic and environmental factors. In the past few decades, many factors, such as smoking, hypertension, diabetes mellitus (DM), arteriosclerosis, obesity and dieting...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/113
CPCC12Q1/6883C12Q2600/118C12Q2600/156C12Q2600/166
Inventor 雷桅陈灿税晓容
Owner 雷桅
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