Individualized-medication gene detection system kit for cardiovascular diseases and application of individualized-medication gene detection system kit for cardiovascular diseases
A gene detection and kit technology, which is applied in the field of genetic detection system kits for individualized drug use in cardiovascular diseases, to achieve accurate results, reduce pollution, and avoid wrong drug use
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Embodiment 1
[0049] Sample collection and DNA extraction
[0050] Collect 200 μl of blood from patients with cardiovascular disease, and use the Axygen DNA extraction kit to extract genomic DNA. For specific methods, refer to the kit instructions. DNA was quality controlled using 0.8% agarose gel electrophoresis and OD260 / 280 values.
[0051] Multiplex PCR
[0052] The multiplex PCR primer sequences of 19 SNP sites related to the metabolic pathway of cardiovascular disease therapeutic drugs use the primer sequences published in the present invention. The 19 pairs of primers were divided into two groups for multiplex PCR amplification. Group 1 includes 10 SNP sites: rs4149056, rs12248560, rs4986893, rs10306114, rs4343, rs5186, rs4244285, rs1057910, rs776746, rs2108622; Group 2 includes 9 SNP sites: rs429358, rs671, rs594918, rs5, 6 rs1801253, rs1065852, rs12041331. The PCR system uses 15μl system: 1.5mM MgCl2, 0.2mMdNTP, 2μl primer mix, 2μl DNA template, 1U Takara hot start Taq enzyme (...
Embodiment 2
[0058] Clinical genetic testing application
[0059] Sample collection and DNA extraction
[0060] 237 blood samples came from patients with cardiovascular diseases in Shanghai Songjiang District Central Hospital, including hypertension, coronary heart disease, chronic heart failure, hyperlipidemia or myocardial infarction. Please see Appendix 1 for patient information. Take 200 μl of blood, and use the Axygen DNA extraction kit to extract genomic DNA. For specific methods, refer to the kit instructions. DNA was quality controlled using 0.8% agarose gel electrophoresis and OD260 / 280 values.
[0061] Table 1 shows the age and gender distribution of patients with blood samples
[0062] information men and women upper age limit lower age limit 50~59(y) 60~69(y) 70~79(y) 80~89(y) 90~100(y) number 106 / 131 98 53 10 105 71 48 3
[0063] method evaluation
[0064] In order to type 19 SNPs at the same time, the probes for LDR detection were de...
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