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Atherosclerosis-related serum miRNA (microribonucleic acid) marker group, and specific primers and application thereof

An atherosclerosis, specific primer technology, applied in the field of medical molecular biology, to achieve the effect of improving sensitivity and specificity, strong complementarity, and accurate detection and quantification

Active Publication Date: 2013-06-19
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, none of the existing early diagnostic markers for atherosclerosis is aimed at the characteristics of serum miRNA expression changes in the disease, and does not involve the combination of miR-135b and miR-499a-3p derived from the changes in the serum miRNA expression profile provided by the present invention as an arterial Application of Molecular Markers in Early Diagnosis of Atherosclerosis

Method used

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  • Atherosclerosis-related serum miRNA (microribonucleic acid) marker group, and specific primers and application thereof
  • Atherosclerosis-related serum miRNA (microribonucleic acid) marker group, and specific primers and application thereof
  • Atherosclerosis-related serum miRNA (microribonucleic acid) marker group, and specific primers and application thereof

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Experimental program
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Embodiment 1

[0029] 1. 54 cases of patients with coronary heart disease and 35 cases of normal healthy controls received physical examination, 5ml of peripheral blood was extracted using coagulation tubes, centrifuged in two steps at 4°C, and finally the upper serum was taken into RNase / DNase-free centrifuge tubes.

[0030] 2. Serum total RNA extraction: TRI Reagent BD was used to extract RNA from serum.

[0031] (1) Lysis: 0.75ml TRI Reagent BD+0.25ml serum;

[0032] (2) Two-phase separation: lysate + 0.1ml bromochloropropane or 0.2ml chloroform;

[0033] (3) RNA precipitation: aqueous phase layer + 0.5ml isopropanol;

[0034] (4) Wash RNA: 1ml 75% ethanol;

[0035] (5) RNA dissolution: dissolve RNA with FORMAzol, water or 0.5% SDS solution prepared by pipetting and mixing with a pipette tip.

[0036] 3. miRNA chip hybridization: using Exiqon miRCURY LNA TM The 11.0 chip includes all miRNAs from the three species of human, mouse, and rat in the miBase database. The capture probes on ...

Embodiment 2

[0047] Using the specific primers of miR-135b and miR-499a-3p (as shown in Table 3), hsa-miR-135b and hsa-miR- There are significant differences in the expression of 499a-3p between the atherosclerosis case group and the healthy control group, such as Figure 4 shown.

[0048] Table 3. Primer information for related miRNAs (Note: GSP is the corresponding specific primer, R is the primer that matches the RT primer)

[0049]

[0050] According to the above results, by analyzing the expression levels of miRNAs in two groups of serum samples (case group and healthy control group), the 90th percentile of the expression levels of miR-135b and miR-499a-3p in the healthy control group was used as the threshold. Grouped by miR-135b and miR-499a-3p, the expression level was less than the 90th percentile, scored as 0, divided into the healthy group, and the expression level was greater than or equal to the 90th percentile, scored as 1 point, divided into the case group, Then evaluat...

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Abstract

The invention relates to an atherosclerosis-related serum miRNA (microribonucleic acid) marker group, and specific primers and application thereof. The marker group comprises a miR-135b marker of which the nucleotide sequence is shown as SEQ ID NO.1 and a miR-499a-3p marker of which the nucleotide sequence is shown as SEQ ID NO.2. The invention also relates to specific primers and application of the miRNA marker group. The two markers screened out from the serum miRNA marker group provided by the invention have high complementarity, thereby ensuring that the detection is quantitatively precise and greatly improving the sensitivity and specificity of disease diagnosis.

Description

technical field [0001] The invention relates to a serum miRNA marker group related to atherosclerosis, specific primers and applications thereof, and belongs to the technical field of medical molecular biology. Background technique [0002] Cardiovascular disease seriously endangers human health. According to WHO statistics, cardiovascular disease has become the first cause of death in the world. The epidemic trend of cardiovascular disease in my country is completely consistent with the global trend. By 2006, it has accounted for 1 / 3 of the total death toll in the country. It is the number one killer of urban and rural residents. Cardiovascular disease has become one of the important factors restricting the social and economic development of our country. Atherosclerosis (AS) is the pathological basis of most cardiovascular diseases and the main cause of disability and death in cardiovascular diseases. The current research consensus is that atherosclerosis biomarkers (Biomar...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/113C12N15/11
Inventor 李曦
Owner SHANDONG UNIV
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