Application of ALKBH2 gene to diagnosis of cerebral ischemic stroke

A technology for ischemic stroke and genes, applied in the direction of recombinant DNA technology, microbiological measurement/testing, DNA/RNA fragments, etc., can solve the problem of not being able to provide specific and sensitive diagnosis, and achieve the effect of timely gene diagnosis

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

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Problems solved by technology

However, the use of the above methods cannot provide a specific and sensitive diagnosis. Although some genes related to ischemic stroke have been reported, th

Method used

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  • Application of ALKBH2 gene to diagnosis of cerebral ischemic stroke
  • Application of ALKBH2 gene to diagnosis of cerebral ischemic stroke
  • Application of ALKBH2 gene to diagnosis of cerebral ischemic stroke

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1 Screening for Gene Markers Related to Ischemic Stroke

[0046] 1. Sample collection

[0047] 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.

[0048] 2. RNA sample preparation and quality analysis

[0049] 2.1 Preparation of RNA samples

[0050] Total RNA was extracted using the RNA extraction kit from Promega. Specific steps are as follows:

[0051] 1) Take 1ml of whole blood collected in a test tube treated with heparin or EDTA, and put it into a sterile centrifuge tube;

[0052] 2) Collect blood cells: centrifuge at 3000rpm (400g) for 5min, carefully suck off the supernatant from the top of the sample;

[0053] 3) Add 1ml of blood cell lysate, pipette carefully 4-5 times, and resuspend the sediment;

[0054] 4) Centrifuge at 3000rpm for 5min;

[0055] 5) Repeat steps 3), 4) twice (three times in total);

[0056] 6) Avoid the...

Embodiment 2

[0081] Example 2 QPCR sequencing verification of differential expression of ALKBH2 gene

[0082]1. According to the detection results of high-throughput sequencing, the ALKBH2 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.

[0083] 2. The RNA extraction steps are the same as in Example 1.

[0084] 3. Reverse transcription: use the reverse transcription kit of TAKARA company to operate. Specific steps are as follows:

[0085] (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;

[0086] (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;

[0087] (3) After 42°C water bath for 60 minutes, 9...

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Abstract

The invention discloses application of ALKBH2 gene to diagnosis of cerebral ischemic stroke. The ALKBH2 gene has low expression in cerebral ischemic stroke, and whether a patient has cerebral ischemic stroke or has a possibility of suffering from cerebral ischemic stroke or not can be judged by detecting the expression level of the ALKBH2 gene. According to the application of ALKBH2 gene to diagnosis of cerebral ischemic stroke, provided by the invention, the diagnostic sensitivity and specificity of cerebral ischemic stroke can be greatly improved, and an early diagnosis means of cerebral ischemic stroke is further provided.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the use of ALKBH2 gene in the diagnosis of 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. There are four types of cerebral ischemia: transient ischemic attack, reversible neurological deficit, progressive stroke, and complete stroke. Ischemic stroke is one of the three major causes of death in the world and the first cause of disability in adults. Since the pathogenesis of ischemic stroke is not yet fully understood, and the means for clinical treatment of ischemic stroke are limited, the disease not only has a high incidence rate, but also has a high mortality rate, disability rate and recurrence rate. [0003] Ischemic stroke is a complex disease with the combined effects of genetic and environmenta...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/68C12N15/11
CPCC12Q1/6883C12Q1/6837C12Q1/6841C12Q1/686G01N33/6803C12Q2561/113C12Q2545/114C12Q2531/113
Inventor 杨承刚邵小健
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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