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SNP marker relevant with treatment of ischemic stroke through antiplatelet drug clopidogrel and application thereof

A technology of ischemic stroke and antiplatelet drugs, which is applied in the field of genetic engineering and cerebrovascular medicine, can solve the problems of different drug reactivity and achieve the effects of improving drug efficacy, improving sensitivity and specificity, and being easy to detect

Active Publication Date: 2017-05-24
SUN YAT SEN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although taking clopidogrel can bring good benefits to most patients with ischemic stroke, more and more studies in recent years have found that different patients have different drug responses to it

Method used

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  • SNP marker relevant with treatment of ischemic stroke through antiplatelet drug clopidogrel and application thereof
  • SNP marker relevant with treatment of ischemic stroke through antiplatelet drug clopidogrel and application thereof
  • SNP marker relevant with treatment of ischemic stroke through antiplatelet drug clopidogrel and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1 Selection and Collection of Research Samples and Arrangement of Sample Data

[0054] 1. Sample selection

[0055] (1) Aged 40-80 years old, gender is not limited;

[0056] (2) Conform to the diagnostic criteria revised by the Fourth National Cerebrovascular Disease Conference and confirmed by CT or MRI;

[0057] (3) Oral clopidogrel at 75mg / day (including the first loading dose of 300mg) for more than 5 days (including 5 days).

[0058] 2. The inventor selected a total of 313 samples that met the criteria for a single Sequenom MassARRAY genotyping experimental sample, and systematically collected the demographic data and clinical data of these samples.

Embodiment 2

[0059] Example 2 Detection of Platelet Aggregation Rate of Sample——Determination of Clopidogrel Resistance

[0060] 1. The platelet aggregation test was carried out by light turbidimetry. Take 2.7 ml of EDTA anticoagulated whole blood (1:9), centrifuge at 800rpm for 5min to extract platelet-rich plasma, then centrifuge at 3000rpm for 15min to extract platelet-poor plasma, the platelet count is 10-20×10 9 / L, use PPP to adjust PRP to 200~300×10 9 / L, PPP was used as blank control, preheated at 37°C for 3 minutes, and 10 μM ADP was used as inducer, and the platelet aggregation rate of patients was measured by light turbidimetry.

[0061] 2. Determination of the platelet aggregation rate in patients with ischemic stroke before taking clopidogrel (day0) and after taking clopidogrel (day5). If the platelet inhibition rate is less than 10% or the blood aggregation value is >50% after taking the drug, it is considered that clopidogrel resistance occurs; according to this standard, ...

Embodiment 3

[0063] Example 3 Genotyping and scanning of SNP markers in peripheral blood DNA

[0064] Among the above-mentioned 313 eligible ischemic stroke patients (including 106 cases in the resistance group and 207 cases in the non-resistance group), relevant results were obtained by Sequenom MassARRAY typing. The specific steps are:

[0065] 1. Extraction of genomic DNA from peripheral blood by phenol-chloroform method

[0066] (1) Take 100 μl of EDTA anticoagulated whole blood, add 200 μl of sterile double distilled water and mix well;

[0067] (2) Add 200 μl of 6 M sodium iodide and mix well;

[0068] (3) Add 400 μl of chloroform / isoamyl alcohol (24:1, v / v), mix repeatedly, let stand for 10 minutes; centrifuge at 12000 rpm for 10 minutes, draw the supernatant and put it in another centrifuge tube;

[0069] (4) Add 300 μl of isopropanol to the supernatant, mix well, let stand for 3 minutes, centrifuge at 12000 rpm for 10 minutes, and discard the supernatant;

[0070] (5) Add 500 ...

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Abstract

The invention discloses a SNP marker relevant with treatment of ischemic stroke through antiplatelet drug clopidogrel and application thereof, as well as a special amplification primer and a special extension primer for detecting the marker. The SNP marker is combination of rs6686001, rs2302429, rs2242480, rs4244285 and rs2046934. Sequences of each marker, the special amplification primer and the special extension primer are sequentially respectively as shown in SEQ ID No. 1-20. According to the SNP marker combination, clopidogrel-resistant and clopidogrel-effective ischemic stroke people can be distinguished well. The SNP marker, the amplification primer thereof and a relevant kit can be used for clinical medication guidance for adopting the antiplatelet drug clopidogrel to treat ischemic stroke patients, medicine resistance is prevented, and a treatment effect is enhanced.

Description

technical field [0001] The invention belongs to the technical fields of genetic engineering and cerebrovascular medicine. More specifically, it relates to a SNP marker associated with the antiplatelet drug clopidogrel in the treatment of ischemic stroke and its application. Background technique [0002] Ischemic stroke, also known as cerebral infarction or cerebral infarction (called stroke in traditional Chinese medicine), refers to the localized ischemic necrosis or softening of the brain. In my country, with the development of society and the continuous changes in residents' lifestyles and eating habits, stroke has become the second leading cause of death in my country, seriously threatening the health of residents. [0003] At present, according to evidence-based medicine, anti-platelet aggregation therapy is the cornerstone of effective prevention and treatment of cerebral infarction while actively intervening in the risk factors of ischemic stroke such as hypertensio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6883C12Q2600/106C12Q2600/156
Inventor 金晶黄民陈易北蔡珮蘅徐春草白旭鹏余伟邦
Owner SUN YAT SEN UNIV
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