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Detection kit and detection method for medication of hypertension patients

A detection kit and detection method technology, applied in the field of genetic detection, can solve the problems of patients who cannot provide medication guidance, low specificity, and no obvious advantages, so as to achieve accurate drug guidance, improve accuracy and specificity Reasonable effect of gender and locus coverage

Inactive Publication Date: 2019-08-16
广州海思医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] (2) The detection site is single, and the evaluation is not accurate enough
75% of hypertensive patients are H-type hypertension, and often accompanied by stroke, myocardial infarction, heart failure, atherosclerosis and other diseases, often need to be combined with aspirin and statins. Medication testing cannot provide comprehensive medication guidance for the corresponding patient population
[0010] (4) There are various detection methods, but none of them have obvious advantages
At present, the products on the market for the guidance of hypertension medication mainly use Sanger method, gene chip method, PCR-melting curve method, fluorescent PCR method, etc. The above methods may have the problems of cumbersome operation and high cost, or have low accuracy and specificity. not strong problem

Method used

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  • Detection kit and detection method for medication of hypertension patients
  • Detection kit and detection method for medication of hypertension patients
  • Detection kit and detection method for medication of hypertension patients

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] The amplification primer probe mainly contains 15 loci of 14 genes. The synthesis of the primers is to design the amplification primers according to the regions of the corresponding sites of the corresponding genes, and the sequences of the primers and the probes are as described in the summary of the present invention. The probe is a TaqMan probe, the fluorescent reporter group of the wild type is FAM, and the fluorescent reporter group of the mutant type is VIC.

Embodiment 2

[0076] The kit includes: detection reaction solution, wild-type quality control, homozygous mutant quality control, heterozygous mutant quality control and blank quality control. Detection reaction solution includes: 10×Buffer, 10×Solution, dNTP, MgSO 4 , primer mix, probe mix, KCl, Taq enzyme, sterilized purified water. Wild-type quality control: wild-type plasmid, homozygous mutant quality control: mutant plasmid, heterozygous mutant quality control: wild-type and mutant plasmid mixture, blank quality control: sterile purified water.

[0077] Specifically in a PCR amplification embodiment, the system used is as follows:

[0078] Component (concentration) Dosage (μL) 10×Buffer 2 10×Solution 2 dNTPs (2.5mM) 1-2 MgSO4(25mM) 0.5-1.5 KCl(1mol / L) 0.5-1.5 Primer mix(10μM) 0.1-0.6 Probe mix(10μM) 0.1-0.5 Taq enzyme (5U / μL) 0.01-0.1 sample DNA 2 Sterilized purified water Add water to 20

[0079] The primer...

Embodiment 3

[0087] The genotype detection of human samples using the above-mentioned kit is as follows:

[0088] 1) Take human peripheral anticoagulated blood, use a commercial DNA extraction kit to process the sample, and extract DNA. The extracted DNA should meet OD260 / OD280=1.8-2.0, concentration>0.5ng / μL;

[0089] 2) Dispense the detection reaction solution into 18 μL / well of the PCR reaction well, and add the DNA of the sample to be tested, the wild-type quality control, the homozygous mutant quality control, the heterozygous mutant quality control and the blank quality control 2 μL each;

[0090] 3) Run according to the following PCR amplification conditions;

[0091]

[0092] 4) Interpretation of results

[0093] Internal standard gene GAPDH: ROX channel Ct ≤ 33, the quality of nucleic acid is qualified, and the results can be analyzed.

[0094] Wild type: FAM channel Ct≤35, corresponding VIC channel Ct>35, the detection result is wild type;

[0095] Homozygous mutant: FAM ...

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Abstract

The invention belongs to the technical field of gene detection, in particular to a detection kit and a detection method for medication of hypertension patients. The medication gene locus combination for hypertension patients totally comprises 15 gene loci, comprising common drug metabolizing enzyme gene loci, drug targeting gene loci and gene loci of common drugs for H-type hypertension and otherhypertension complications. The gene loci are based on a fluorescence quantitative PCR technology and uses a specific Taqman primer probe set to greatly improve the accuracy and specificity of the product; the locus coverage is reasonable, the guiding effect on drugs is more accurate, a comprehensive reference is provided for the combined medication and individualized medication of hypertension patients, guidance can be provided for people suffering from hypertension and complications thereof, patients can receive the most effective treatment as soon as possible, and the whole detection process only needs 4 hours from extraction to result determination.

Description

technical field [0001] The invention belongs to the technical field of gene detection, and in particular relates to a detection kit and a detection method for patients with hypertension. Background technique [0002] According to the "Report on Nutrition and Chronic Disease Status of Chinese Residents (2015)", the prevalence of hypertension in Chinese residents aged ≥18 was 25.2% in 2012, and the number of hypertensive patients in China was 270 million. Hypertension is also the most important risk factor for cardiovascular and cerebrovascular diseases, with major complications such as stroke, myocardial infarction, heart failure, atherosclerosis, and chronic kidney disease. [0003] Drug therapy is currently the most important measure to control blood pressure. The main clinical antihypertensive drugs are diuretics, calcium channel blockers (CCB), angiotensin converting enzyme inhibitors (ACEI), β-blockers, and angiotensin II receptors. There are five categories of angiote...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12Q1/6858
CPCC12Q1/6858C12Q1/6883C12Q2600/106C12Q2600/156C12Q2531/113C12Q2561/101C12Q2545/101
Inventor 夏昊强周煌凯王芳刘倩陆嫚云曹治家姚啟聪李洁莹
Owner 广州海思医疗科技有限公司
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