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High-throughput sequencing method for hypertension treatment drug related genes and application thereof

A technology for the treatment of drugs and hypertension, applied in the determination/examination of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., which can solve the problems of limited drug types, low flexibility, and low throughput

Pending Publication Date: 2019-11-29
HANGZHOU HEYI GENE TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The real-time fluorescent PCR method can only detect a few common mutation sites, and the types of drugs covered are limited, and the method has low throughput and high cost
Although the gene chip method can detect multiple known SNP sites to be tested at the same time, it is expensive, has low flexibility, and can only detect known sites

Method used

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  • High-throughput sequencing method for hypertension treatment drug related genes and application thereof
  • High-throughput sequencing method for hypertension treatment drug related genes and application thereof
  • High-throughput sequencing method for hypertension treatment drug related genes and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] Detection of drug-related genes for hypertension treatment based on Illumina sequencing platform

[0078] Reagents: NEBNext Ultra II Q5 Master Mix (New England Biolabs), Agencourt AMPureXP (Beckman Coulter), QubitdsDNA HS Assay Kit (ThermoFisher), MagMAX TM DNA Multi-Sample Kit (ThermoFisher), primers for gene amplification related to antihypertensive drugs (one gene), I5 end adapter primer (one gene), I7 end adapter primer (one gene)

[0079] 1. Genomic DNA extraction and concentration detection of the sample to be tested:

[0080] Take three hypertensive patients: sample 1, sample 2, sample 3 anticoagulant blood samples, use MagMAX TM DNA Multi-Sample Kit, follow the kit instructions for DNA extraction. For the extracted genomic DNA, the concentration of the sample was quantitatively detected by Qubit. The result is as follows:

[0081] sample sample 1 sample 2 sample 3 Concentration (ng / μL) 46 39 25

[0082] 2. Multiplex PCR amplifi...

Embodiment 2

[0126] Three hypertensive patients were taken: sample 1, sample 2, and sample 3 anticoagulant blood samples were tested, and the operation steps were operated according to Example 1. The following results were obtained after sequencing on the Illumina sequencing platform and data analysis:

[0127]

[0128]

[0129] Losartan is an angiotensin II receptor antagonist and is a commonly used antihypertensive drug. The protein encoded by CYP2C9 belongs to the CYP450 family of drug-metabolizing enzymes, which can catalyze the metabolism of losartan into the active product EXP-3174 in vivo, thereby exerting the antihypertensive effect. Some polymorphisms of CYP2C9 gene affect the production of losartan active product EXP-3174, and then affect the antihypertensive effect. Individuals carrying the CYP2C9*3 (rs1057910, A1075C, Ile359Leu) allele had reduced production of EXP-3174 and reduced metabolic rate of losartan after administration of losartan. Oral administration of a sing...

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Abstract

The invention belongs to the field of hypertension treatment drug related gene detection, and particularly relates to a high-throughput sequencing method for hypertension treatment drug related genesand an application of the high-throughput sequencing method. The invention discloses the high-throughput sequencing method for hypertension treatment drug related genes and the application of the high-throughput sequencing method. The method comprises the following steps: determining hypertension treatment drug related genes, designing a hypertension treatment drug related gene amplification primer combination, extracting genome DNA and detecting concentration, performing multiple a PCR amplification reaction and product purification on the hypertension treatment drug related gene, performingjoint sequence PCR reaction and product purification, performing library quality inspection, performing high-throughput sequencing and the like. The method can realize typing detection of 39 SNP lociof 33 hypertension treatment drug related genes at one time, 27 common hypertension treatment medicines are covered, more detection sites, higher flux, higher data accuracy and higher economy are realized, guidance can be provided for individualized medication of patients, the medicine treatment efficiency is improved, and the risk of toxic and side effects is reduced.

Description

technical field [0001] The invention belongs to the field of detection of genes related to drugs for treating hypertension, and in particular relates to a high-throughput sequencing method for genes related to drugs for treating hypertension and its application. Background technique [0002] Hypertension is one of the most common chronic diseases in my country. In the 2019 "Guidelines for the Prevention and Treatment of Hypertension in China 2018 Revised Edition", the latest survey data on hypertensive disorders in my country shows that from 2012 to 2015, the number of hypertensive patients aged 18 and above in my country The crude disease rate is 27.9%, and the prevalence rate of hypertension in our country is still on the rise. As one of the most important risk factors for cardiovascular and cerebrovascular diseases, hypertension has a high disability and mortality rate and a serious epidemic situation. It has caused heavy economic and spiritual burdens on families and soci...

Claims

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Application Information

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IPC IPC(8): C12Q1/6869C12Q1/6883C12N15/11
CPCC12Q1/6869C12Q1/6883C12Q2600/16C12Q2600/106C12Q2600/156C12Q2525/191C12Q2531/113C12Q2537/143C12Q2565/125C12Q2535/122
Inventor 王军一肖雯闫楠王晓娟叶可勇舒平王兆宝高金龙
Owner HANGZHOU HEYI GENE TECH
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