Molecular marker for diagnosing cerebral ischemic thrombosis

A technology of ischemic stroke and molecular markers, applied in disease diagnosis, analytical materials, biological testing, etc., can solve problems such as increasing the risk of stroke

Inactive Publication Date: 2016-09-07
BEIJING MEDINTELL BIOMED CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] At present, stroke is considered to be a complex disease affected by environmental factors, genetic factors, etc. Although the pathogenesis of stroke has not been fully understood, hypertension, diabetes, dyslipidemia, obesity, smoking and excessive drinking have been It is recognized as the main independent risk factor for stroke, and the aggregation of these risk factors will further increase the risk of stroke

Method used

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  • Molecular marker for diagnosing cerebral ischemic thrombosis
  • Molecular marker for diagnosing cerebral ischemic thrombosis
  • Molecular marker for diagnosing cerebral ischemic thrombosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 Screening for Gene Markers Related to Ischemic Stroke

[0044] 1. Sample collection

[0045] 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.

[0046] 2. Extraction and quality analysis of sample total RNA

[0047] 2.1 Extraction of sample RNA

[0048] Total RNA was extracted using the Blood RNA extraction kit from U-gene. Specific steps are as follows:

[0049] 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;

[0050] 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;

[0051] 3) Centrifuge at 450g for 10min at 4°C to precipit...

Embodiment 2

[0079] Example 2 QPCR sequencing to verify the differential expression of CTAGE5 gene

[0080] 1. According to the detection results of high-throughput sequencing, the CTAGE5 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.

[0081] 2. The RNA extraction steps are the same as in Example 1.

[0082] 3. Reverse transcription: use the reverse transcription kit of TAKARA company to operate. Specific steps are as follows:

[0083] (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;

[0084] (2) Construct a 25 μl reaction system, including 5 μl of 5× reverse transcription buffer, 5 μl of dNTP (2.5 mM), 40 U / μl of RNasin, 200 U / μl of M-MLV, and make up to 25 μl of nuclease-free water;

[0085] (3) After 60 min in water bath at 42°C...

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Abstract

The invention discloses a molecular marker for diagnosing cerebral ischemic thrombosis. The molecular marker is a CTAGE5 gene, and early diagnosis can be provided for a patient with the cerebral ischemic thrombosis by detecting expression level of the CTAGE5 gene, so that early detection and early treatment are realized, and death rate of the patient with the cerebral ischemic thrombosis is reduced. The invention provides a cerebral ischemic thrombosis diagnosis product. The product comprises a reagent for detecting the expression level of the CTAGE5 gene and can realize the early diagnosis on the cerebral ischemic thrombosis.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a molecular marker for diagnosing ischemic stroke, specifically the molecular marker is CTAGE5. Background technique [0002] Stroke is a refractory disease that seriously endangers the safety of human life and is characterized by high morbidity, disability and mortality. In the ranking of the causes of death published by the Ministry of Health of my country in 2008, stroke is the third leading cause of death in urban areas and the second leading cause of death in rural areas. Epidemiological surveys show that the incidence of stroke in young people has been increasing in recent years. The proportion of stroke in adults under the age of 45 is 5% and 13.44% in Western countries and domestic reports, respectively. Stroke is divided into ischemic stroke and hemorrhagic stroke according to its nature, and ischemic stroke accounts for 75% to 85% of the total number of stroke patients. Isc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/68G01N33/53
CPCC12Q1/6883C12Q2600/158G01N33/53G01N33/6893G01N2333/47G01N2800/2871
Inventor 宋宏涛李曙光杨承刚
Owner BEIJING MEDINTELL BIOMED CO LTD
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