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Gene marker related to ischemic stroke

An ischemic stroke and genetic technology, applied in the biological field, to achieve the effect of reducing mortality, timely genetic diagnosis, and more accurate genetic diagnosis

Active Publication Date: 2016-02-03
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the diagnosis of stroke mainly relies on clinical examination and various neuroimaging techniques. Although medical technology has been highly developed compared with the past, there are still no reliable biomarkers that can predict the risk and early diagnosis of ischemic stroke. diagnosis

Method used

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  • Gene marker related to ischemic stroke
  • Gene marker related to ischemic stroke
  • Gene marker related to ischemic stroke

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 Screening for Gene Markers Related to Ischemic Stroke

[0047] 1. Sample collection

[0048] 10 normal blood samples and blood samples from patients with ischemic stroke were collected, and all the above samples were obtained with the consent of the ethics committee.

[0049] 2. RNA sample preparation and quality analysis

[0050] 2.1 Preparation of RNA samples

[0051] (1) Homogenization treatment

[0052] Take blood directly, add 3 times the volume of erythrocyte lysate, mix well, place at room temperature for 10 minutes, and centrifuge at 10,000 rpm for 1 minute. Discard the supernatant thoroughly and collect the white blood cell pellet. Add 1ml Trizol per 100-200μl blood collected leukocyte pellet.

[0053] (2) layered

[0054] a. After adding TRIzol to the sample, place it at room temperature for 5 minutes to fully lyse the sample. Centrifuge at 12,000rpm at 4°C for 10min, and take the supernatant;

[0055] b. Add 200 μl of chloroform to every 1ml...

Embodiment 2

[0077] Example 2 QPCR sequencing to verify the differential expression of the METTL9 gene

[0078] 1. According to the detection results of high-throughput sequencing, the METTL9 gene was selected for large-sample QPCR verification. According to the sample collection method in Example 1, 80 cases of ischemic stroke patients' blood and 80 normal people's blood were selected.

[0079] 2. The RNA extraction steps are the same as in Example 1.

[0080] 3. Reverse transcription: use the reverse transcription kit of TAKARA company to operate. Specific steps are as follows:

[0081] (1) Take 2 μg of total RNA for reverse transcription, add 2 μl of Oligo(dT) and mix well; 70°C water bath; 5 min and then immediately ice bath for 2-3 min;

[0082] (2) Construct a 25 μl reaction system, including 5 μl of 5× reverse transcription buffer, 5 μl of dNTP (2.5 mM), RNasin 40 U / μl, M-MLV 200 U / μl, and supplement nuclease-free water to 25 μl;

[0083] (3) After 60 min in water bath at 42°C, ...

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Abstract

The invention discloses a gene marker related to ischemic stroke. The expression of an METTL9 gene in ischemic stroke is upregulated, and early diagnosis can be provided for ischemic stroke patients by detecting the expression level of the METTL9 gene, so that early detection and treatment can be realized to reduce mortality of the ischemic stroke patients.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a gene marker METTL9 related to ischemic stroke. Background technique [0002] Ischemic stroke, also known as cerebral infarction, refers to ischemic necrosis or encephalomalacia of limited brain tissue caused by cerebral blood supply disturbance, ischemia and hypoxia. Mechanisms such as energy metabolism disorder, excitatory amino acid toxicity, inflammatory response, penumbra depolarization, and cell apoptosis are all involved in its pathophysiological process. There are generally four types of cerebral ischemia: transient ischemic attack, reversible neurological deficit, progressive stroke, and complete stroke. Ischemic stroke is a disease that seriously endangers human health, is one of the three major diseases that cause human death, and is the main cause of limb paralysis. , The recurrence rate of ischemic stroke is high, as high as 30% in domestic reports. [0003] With the...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/68
CPCC12Q1/6883C12Q2600/158G01N33/6893G01N2800/2871G01N2800/50
Inventor 董琳杨承刚
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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