Cerebral-arterial-thrombosis miRNA marker

An ischemic stroke, miRNA-4252 technology, applied in the field of biomedicine, can solve problems such as abnormal gene expression and induced diseases

Active Publication Date: 2016-02-24
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The abnormal expression of miRNA directly leads to the abnormal expression of some genes related to the occurrence of diseases, which induces the occurrence of diseases.
Although it has been reported that miRNA can play an i...

Method used

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  • Cerebral-arterial-thrombosis miRNA marker
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  • Cerebral-arterial-thrombosis miRNA marker

Examples

Experimental program
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Effect test

Embodiment 1

[0035] Example 1 Screening of miRNAs associated with ischemic stroke

[0036] 1. Sample acquisition: Collect blood samples from 10 healthy people and ischemic stroke patients. All the above specimens were obtained with the consent of the organizational ethics committee.

[0037] 2. Extraction of total RNA from samples

[0038] Total RNA was extracted using the BloodRNA extraction kit from U-gene. Specific steps are as follows:

[0039] 1) Add 5 times the volume of 1×XR-I buffer per volume of fresh blood (maximum 1ml), for example: add 5ml of XR-I buffer per 1ml of blood, and vortex to mix;

[0040] 2) Cool in ice for 15 minutes, and quickly mix twice on a vortex shaker. The solution becomes clear, indicating that the red blood cells have been lysed. If the hematocrit or ECR of individual samples increases, extend the ice bath time to 20 minutes;

[0041] 3) Centrifuge at 450g for 10min at 4°C to precipitate white blood cells, and completely discard the supernatant contain...

Embodiment 2

[0067] Example 2 QPCR verification of differentially expressed miRNA-4252

[0068] 1. According to the detection results of the miRNA chip, select miRNA-4252 for large-sample QPCR verification. According to the sample collection method in Example 1, 80 blood samples from patients with ischemic stroke and 80 blood samples from healthy people were selected.

[0069] 2. The RNA extraction process is the same as in Example 1.

[0070] 3. Reverse transcription:

[0071] 1) Mix 10 pg-1 μg of total RNA template with 2 μl 10× buffer, 2 μl ldATP (10 mM), 0.5 μl polyA polymerase, 0.5 μl ribonuclease (RNase) inhibitor and ribonuclease-free water (RNasefreewater), the final volume is 20 μl , and incubated at 37°C for 1h.

[0072] 2) Add 1 μl of 0.5 μg / μl Oligo(dT) specific RT primer to the reaction tube, and incubate at 70° C. for 5 minutes.

[0073] 3) Immediately incubate on ice for at least 2 minutes to break the secondary structure of RNA and primers.

[0074] 4) Mix the above 20...

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Abstract

The invention discloses a miRNA marker. The miRNA marker is miRNA-4252. Whether a patient suffers from cerebral arterial thrombosis or not can be diagnosed by detecting the expression level of the miRNA-4252. An experiment shows that a cerebral arterial thrombosis patient and a healthy person can be effectively differentiated through the miRNA-4252. On the basis, the miRNA-4252 can be used for preparing a product for diagnosing the cerebral arterial thrombosis. By means of the cerebral-arterial-thrombosis miRNA marker, the sensitivity and the specificity of diagnosing of the cerebral arterial thrombosis can be greatly improved, and early diagnosis of the cerebral arterial thrombosis is achieved.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to an ischemic stroke miRNA marker and application thereof. Background technique [0002] Stroke is a group of diseases that cause brain tissue damage due to sudden rupture of blood vessels in the brain or blood circulation disturbance caused by vascular obstruction, including ischemic stroke and hemorrhagic stroke. Ischemic stroke refers to local brain tissue including Necrosis of nerve cells, coelomocytes, and blood vessels due to lack of blood supply. There are about 1.5 to 2 million new stroke patients in my country every year, of which ischemic stroke accounts for 60 to 80% of stroke cases. Ischemic stroke can be caused by occlusion and stenosis of the internal carotid artery and vertebral artery. Over 40 years old, and more men than women, severe cases can cause death. [0003] miRNA is a 21-22nt non-coding RNA molecule that naturally exists in the body. It is a class of RNA that reg...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6883C12Q2600/158C12Q2600/178
Inventor 杨承刚孙耀兰
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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