Method and reagent kit for forecasting susceptibility of incidence of intracerebral haemorrhage

A technology for hemorrhagic stroke and susceptibility, applied in the fields of medical biotechnology and genetic diagnosis, can solve the problems of long time, poor repeatability, low sensitivity, etc., and achieve the effects of simple operation, stable results, high sensitivity and accuracy

Inactive Publication Date: 2008-08-20
FUWAI HOSPITAL OF CARDIOVASCULAR DESEASE CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Due to the traditional PCR-enzyme digestion method for SNP analysis, there are problems such as long time, poor repeatability, low sensitivity, and low accuracy.

Method used

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  • Method and reagent kit for forecasting susceptibility of incidence of intracerebral haemorrhage
  • Method and reagent kit for forecasting susceptibility of incidence of intracerebral haemorrhage
  • Method and reagent kit for forecasting susceptibility of incidence of intracerebral haemorrhage

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: Kit for detecting risk associated with hemorrhagic stroke

[0030] 1. Kit components

[0031] The kit for detecting the risk of hemorrhagic stroke contains a pair of primers that can amplify the SNP3379 site of the ANGPT1 gene and corresponding reagents for LDR-PCR. The components and contents are as follows, and stored at -20°C:

[0032] Table 1: PCR amplification reagents (10 persons):

[0033]

[0034]

[0035] 10×HotStar PCR buffer: 100mM Tris-HCl (pH8.3), 500mM KCl, 15mM MgCl 2 , 100 μg / ml BSA.

[0036] 10mM dNTP mixture: 2.5mM each of dATP, dCTP, dTTP, and dGTP.

[0037] The F primer sequence and the R primer sequence are the base sequences shown in SEQ ID NO.2 and SEQ ID NO.3 in the sequence table, respectively.

[0038] Table 2: LDR detection reagents (10 servings)

[0039]

[0040] The LDR probe mixture is the base shown in the sequence table SEQ ID NO.4 to SEQ ID NO.6, wherein the 5' end of SEQ ID...

Embodiment 2

[0068] Example 2: Collection and extraction of genomic DNA

[0069] The samples were collected from the 973 project undertaken by the Sino-German Laboratory of Fuwai Cardiovascular Disease Hospital, Chinese Academy of Medical Sciences. A total of 489 patients with hemorrhagic stroke were collected, with an average age of 58.0±9.7 years, of which 63.2% were male, and 1843 cases of controls, with an average age of 59.3±8.5 years, of which 57.6% were male. All subjects were of Han nationality and signed informed consent, and this study was also approved by the ethics committee.

[0070] Genomic DNA was prepared from human peripheral blood according to the following method:

[0071] In the presence of anticoagulant EDTA, 10 ml of human peripheral blood collected was centrifuged at 2500 rpm for 30 minutes to remove serum. A 0.2% NaCl solution was then added to bring the total volume to 50 ml. The solution was shaken gently 5-6 times and allowed to sit on ice for 15 minutes. The...

Embodiment 3

[0072] Example 3: Correlation study between ANGPT1 gene SNP and hemorrhagic stroke

[0073] 1. Identification and determination of SNP

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Abstract

The invention discloses a method for forecasting hemorrhagic stroke susceptibility, which pertains to the medical biotechnology and the genetic diagnosis field. The method for forecasting the hemorrhagic stroke susceptibility determines the genotype of ANGPT1 gene polymorphism 3379 site (rs2507800) of a subject and forecasts the hemorrhagic stroke susceptibility of the subject by extracting the host cell genome DNA; when the ANGPT1 genotype is AA homozygous, the susceptibility of the subject is highest; and the risk of increasing the hemorrhagic stroke is 1.96 times. The single nucleotide polymorphism (SNP) of the ANGPT1 gene of the invention has the nucleic acid sequences site which is shown in SEQ ID NO.1, and is one of the high-risk sites that are related to the incidence of the hemorrhagic stroke. The method has the advantages that: the relevance of the ANGPT1 gene polymorphism site with hemorrhagic stroke is firstly expounded, the method for forecasting hemorrhagic stroke susceptibility is provided, and the method can be used for the prevention, the auxiliary diagnosis and treatment of hemorrhagic stroke, as well as the development of new drugs.

Description

technical field [0001] The present invention relates to a method and a kit for predicting the susceptibility of hemorrhagic stroke, more specifically, predicting the subject by measuring the polymorphic site 3379A / T of human angiopoietin-1 (angiopoietin 1; ANGPT1) For the susceptibility of hemorrhagic stroke, the method can be used for auxiliary diagnosis, treatment and new drug development of the disease, and belongs to the field of medical biotechnology and gene diagnosis. Background technique [0002] my country is a big country of stroke in the world. In 2003, 57.02 million people died in the world every year, about 1% of the mortality rate. 16.73 million died of cardiovascular and cerebrovascular diseases, accounting for about 1 / 3 of all deaths; among cardiovascular and cerebrovascular diseases, 5.5 million died of stroke, accounting for about 1 / 3; 4.2 million of them were in developing countries, about half of them (2 million) are in China. The incidence rate of ini...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
Inventor 惠汝太杨涛陈敬洲石毅孙凯张禅那
Owner FUWAI HOSPITAL OF CARDIOVASCULAR DESEASE CHINESE ACAD OF MEDICAL SCI
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