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Cerebral-arterial-thrombosis diagnosis marker

A technology of ischemic stroke, miRNA-4511, applied in the field of biomedicine

Inactive 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

However, there is currently no quick and easy laboratory blood test for the screening and early detection of ischemic stroke

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 Screening of miRNAs associated with ischemic stroke

[0039] 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.

[0040] 2. Extraction of total RNA from samples

[0041] Total RNA was extracted using the BloodRNA extraction kit from U-gene. Specific steps are as follows:

[0042] 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;

[0043] 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;

[0044] 3) Centrifuge at 450g for 10min at 4°C to precipitate white blood cells, and completely discard the supernatant contain...

Embodiment 2

[0070] Example 2 QPCR verification of differentially expressed miRNA-4511

[0071] 1. Select miRNA-4511 according to the detection results of the miRNA chip for large sample QPCR verification. According to the sample collection method in Example 1, 70 blood samples from patients with ischemic stroke and 70 blood samples from healthy people were selected.

[0072] 2. The RNA extraction process is the same as in Example 1.

[0073] 3. Reverse transcription: 2 μg of total RNA template was mixed with 1 μl of 2 μM miRNA reverse transcription primer, 1 μl of 2 μM U6snRNA reverse transcription primer, 1 μl of ldNTP (10 mM) mixture and RNase-free deionized water in a final volume of 20 μl. After mixing, incubate at 65°C for 5min. Then immediately cool on ice. Then add the following components in sequence: 4 μl 5× buffer, 2 μl DTT (0.1 M), 1 μl ribonuclease (RNase) inhibitor, mix the solution, and incubate at 37° C. for 5 min. Then add 1 μl of M-MLV reverse transcriptase, mix well ...

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Abstract

The invention discloses a cerebral-arterial-thrombosis diagnosis marker. The marker is miRNA-4511. The miRNA-4511 is highly expressed in a cerebral arterial thrombosis patient, and whether a patient suffers from cerebral arterial thrombosis or not or whether a patient has the risk of suffering from the cerebral arterial thrombosis or not can be diagnosed by detecting the expression level of the miRNA-4511. On the basis, the miRNA-4511 can be used for preparing a product for diagnosing the cerebral arterial thrombosis. As the cerebral-arterial-thrombosis diagnosis marker is discovered, the specificity and the sensitivity of diagnosing of the cerebral arterial thrombosis can be improved, and early diagnosis of the disease is achieved.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a diagnostic marker for ischemic stroke, in particular to a miRNA marker for ischemic stroke. Background technique [0002] Ischemic stroke refers to the focal neurological deficit syndrome caused by the sudden onset of local blood circulation disorder in the brain leading to cerebral ischemia and hypoxia. Ischemic stroke is one of the three major causes of death in the world and the first cause of disability in adults. Due to the fact that the pathogenesis of ischemic stroke has not been fully elucidated, and the ideal clinical treatment methods are limited, not only the morbidity rate is high, but the mortality rate, disability rate and recurrence rate are also high, causing huge harm to individuals and society. burden. [0003] Ischemic stroke is a complex disease characterized by the interaction of genetic and environmental factors. With the completion of the Human Genome Project an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6883C12Q2600/158C12Q2600/178
Inventor 杨承刚边洋
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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