Human cytochrome CYP2C19 and ABCB1 gene polymorphism site detection kit and application thereof

A CYP2C19, gene locus technology, applied in the field of biomedicine, can solve the problems of increased risk of embolism re-formation, increased risk of cardiovascular and cerebrovascular events, and increased mortality

Active Publication Date: 2019-03-05
BEIJING TIANTAN HOSPITAL AFFILIATED TO CAPITAL MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The latest clinical research results confirm that individuals with poor metabolism caused by CYP2C19 gene mutations have an increa

Method used

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  • Human cytochrome CYP2C19 and ABCB1 gene polymorphism site detection kit and application thereof
  • Human cytochrome CYP2C19 and ABCB1 gene polymorphism site detection kit and application thereof
  • Human cytochrome CYP2C19 and ABCB1 gene polymorphism site detection kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0202] Example 1 Establishment of real-time fluorescent quantitative allele-specific PCR and its clinical evaluation and application

[0203] 1. Establishment of real-time fluorescence quantitative allele-specific PCR method

[0204] Real-time fluorescence quantitative PCR technology has the characteristics of real-time monitoring, quantification and high throughput, and is easy to operate and high in sensitivity. Fluorescent quantitative PCR is divided into probe quantitative PCR (Taqman method) and fluorescent dye quantitative PCR. Probe PCR (Taqman method) refers to adding a specific fluorescent probe while adding a pair of primers during amplification. The probe is an oligonucleotide, and the two ends are respectively labeled with a reporter fluorescent group and a quencher. inactivate the fluorophore. When the probe is intact, the fluorescent signal emitted by the reporter group is absorbed by the quencher group; during PCR amplification, the 5'-3' exonuclease activity ...

Embodiment 2

[0251] Example 2: Detection of CYP2C19*2; CYP2C19*3; CYP2C19*17 and ABCB1 Genotypes by Sequencing

[0252] The current gold standard for gene polymorphism is the sequencing method. In this example, the sequencing method is used to study the distribution of CYP2C19*2; CYP2C19*3; CYP2C19*17 and ABCB1 genes in healthy people. The reliability and accuracy of the method of the present invention are evaluated by comparing with the results obtained by real-time fluorescent quantitative PCR Taqman.

[0253] The sequencing method refers to amplifying the target gene fragment of the sample to be tested by conventional PCR technology, performing sequence determination by the traditional dideoxy method, and comparing with the original sequence, so as to analyze the polymorphic site, which is currently the most common method. Methods used for SNP studies. This method can be used to determine the differences at the molecular level of species at different levels within and between populatio...

Embodiment 3

[0299] This embodiment provides a human cytochrome P450 CYP2C19*2; CYP2C19*3; CYP2C19*17; and ABCB1:rs4148727 genomic single nucleotide polymorphism (SNP) real-time fluorescent quantitative PCR Taqman detection kit.

[0300] This kit can identify and detect CYP2C19*2; CYP2C19*3; CYP2C19*17; and ABCB1:rs4148727 loci in the human genome, and is suitable for the typing of this combined SNP analysis. The kit adopts the principle of fluorescent probe hydrolysis method (Taqman method), including the primer concentration and probe concentration most suitable for real-time fluorescent quantitative PCR reaction, and adopts hot-start Taq DNA polymerase and is most compatible with multiple real-time fluorescent quantitative reactions The unique buffer and primer combination can effectively inhibit non-specific PCR amplification and achieve the purpose of high sensitivity and high throughput real-time fluorescence quantitative PCR amplification reaction. When conducting experiments, the p...

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Abstract

The invention provides a human cytochrome CYP2C19 and ABCB1 gene polymorphism site detection kit and application thereof, primers and probes are synthesized according to nucleotide sequences of four polymorphic sites CYP2C19*2: rs4244285; CYP2C19*3: rs4986893; CYP2C19*17: rs12248560 and ABCB1: rs4148727 of two genes of cytochrome P450, and by real-time fluorescence quantitative allele probe detection, whether a target is a drug susceptible population is determined. Using the method, the guidance and adjustment of a clinical drug regimen can be carried out to provide a basis for clinical personalized treatment and prevent adverse drug reactions. The method can simultaneously detect the four polymorphic sites of the two genes, and has the advantages of simplicity, accuracy, rapidity and highthroughput.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a detection kit for human cytochrome P450 CYP2C19 and ABCB1 gene polymorphism sites and its use. Background technique [0002] Ischemic heart disease and ischemic stroke are the top two global causes of death announced by the World Health Organization (WHO). Atherosclerosis, plaque rupture, and thrombosis are the direct causes of cardiovascular and cerebrovascular events, and thrombosis Diseases have become the number one killer of human health, and antiplatelet therapy is the main means of preventing thrombotic diseases. [0003] Plaque rupture and thrombosis are the basic pathological changes leading to acute cardiovascular events, and antithrombotic therapy, especially antiplatelet therapy, is the most important intervention for acute coronary syndrome. Platelets are nucleated, cytoplasmic fragments produced from megakaryocytes in the bone marrow. Its maximum lifetime in circulati...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883
CPCC12Q1/6883C12Q2600/106C12Q2600/156Y02A50/30
Inventor 王拥军王伊龙赵性泉林金嬉李昊林毅潘岳松刘丽萍
Owner BEIJING TIANTAN HOSPITAL AFFILIATED TO CAPITAL MEDICAL UNIV
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