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Circulating miRNA marker for diagnosis of acute myocardial infarction and application of marker

A technology for acute myocardial infarction and myocardial infarction, which is applied in the field of medical molecular biology to achieve the effect of improving sensitivity and specificity

Inactive Publication Date: 2019-07-26
QINGDAO UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] However, up to now, there are few reports on the use of circulating miRNAs in the diagnosis of early acute myocardial infarction, and none of the existing miRNA diagnostic markers for acute myocardial infarction has studied the characteristics of miRNA expression changes before and after percutaneous coronary intervention (PCI).

Method used

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  • Circulating miRNA marker for diagnosis of acute myocardial infarction and application of marker
  • Circulating miRNA marker for diagnosis of acute myocardial infarction and application of marker
  • Circulating miRNA marker for diagnosis of acute myocardial infarction and application of marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0087] Embodiment 1 Sample processing

[0088] 1. 31 patients with acute myocardial infarction and 27 normal healthy controls received physical examination. 5 mL of peripheral blood was drawn from anticoagulant blood vessels, centrifuged in two steps at 4°C, and finally the upper plasma was collected in RNase / DNase-free centrifuge tubes.

[0089] 2. Extraction of plasma total RNA: Use TRI Reagent to extract RNA from plasma:

[0090] (1) Lysis: 0.75 mL TRI Reagent+0.25 mL serum, vigorously mix;

[0091] (2) Internal reference: 50 pM synthetic nematode cel-mir-39 was added to the lysate to provide an internal control for validation studies;

[0092] (3) Two-phase separation: the above lysate + 0.2 mL chloroform, vigorously mixed, let stand for 10 min, and centrifuge at 12000 g for 10 min at 4 °C;

[0093] (4) RNA precipitation: aqueous layer + 0.6 mL isopropanol + 3 μL glycogen, precipitate for several hours;

[0094] (5) Wash RNA: 1 mL of 75% ethanol, centrifuge at 12000 g...

Embodiment 2

[0096] Example 2 Real-time PCR detection

[0097]Use Takara's miRNA detection kit (including reverse transcription kit, specific miRNA primers and probes, fluorescence quantitative detection kit, etc.) to detect miR-26a-1, miR-146a and miR in serum -199a-1 and the level of the reference miRNA (cel-mir-39), the average of the three results (Ct value) is used as the Ct value of the specific miRNA.

Embodiment 3

[0098] The standardized processing of embodiment 3 data

[0099] According to the measured Ct values ​​of miR-26a-1, miR-146a, miR-199a-1 and reference miRNA (cel-mir-39) in the sample serum. With cel-mir-39 as a reference, the relative content of specific miRNA in serum was obtained, and the classic 2 in PCR detection －ΔCt The method of expresses the level of specific miRNA in serum (ΔCt is the difference between the Ct value of target miRNA and reference cel-mir-39). The result is as Figure 1 to Figure 3 shown. From figure 1 In a, it can be seen that the expression level of circulating miR-26a-1 in patients with acute myocardial infarction before and after PCI is significantly higher than that in healthy controls, indicating that miR-26a-1 can be developed as a diagnostic marker. From figure 1 In b, it can be seen that the expression level of circulating miR-146a in patients with acute myocardial infarction before and after PCI is significantly higher than that in heal...

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Abstract

The invention provides an application of an miRNA marker for diagnosing acute myocardial infarction, a product using the miRNA marker and a detection method, and relates to the technical field of medical molecular biology. The miR-26a-1, miR-146a or miR-199a-1 can be singly or jointly applied as markers for assisting early diagnosis of myocardial infarction, and the plasma level of the miR-26a-1,miR-146a or miR-199a-1 is obviously increased in the early stage of occurrence of myocardial infarction; and the miRNA in the blood of patients with acute myocardial infarction is released into the blood earlier than intracellular protein markers, so that the detection of the plasma level of the miRNA can be used as a biomarker for assisting early diagnosis of acute myocardial infarction. The three miRNA markers can be used as markers for early diagnosis of heart diseases, and thus the sensitivity and specificity of disease diagnosis are greatly improved.

Description

technical field [0001] The invention relates to the technical field of medical molecular biology, in particular to an application of miRNA, a product using the same and a detection method. Background technique [0002] Acute myocardial infarction (AMI) is acute myocardial ischemia and hypoxia caused by coronary artery stenosis and blockage, leading to myocardial necrosis. It is one of the main causes of disease mortality and has seriously endangered human health all over the world. The incidence of acute myocardial infarction in my country is about 55 / 100,000, and it is on the rise in recent years. The occurrence of acute myocardial infarction involves a variety of pathological and physiological factors. It is a complex and critical disease with high mortality and disability rates. The current commonly used treatment for acute myocardial infarction is thrombolysis or early vascular stent intervention to restore blood circulation, which has greatly saved patients with acute ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883
CPCC12Q1/6883C12Q2600/158C12Q2600/178
Inventor 薛升朱文杰
Owner QINGDAO UNIV
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