Human cytochrome P450CYP2C19 and SVIL (Supervillin) gene polymorphic site detection kit and application

A CYP2C19, 1.SVIL technology, applied in genetic engineering, plant gene improvement, application, etc., can solve problems such as increased risk of embolism re-formation, increased risk of cardiovascular and cerebrovascular events, and increased mortality

Pending Publication Date: 2017-12-29
龙凌云 +1
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  • Summary
  • 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 P450CYP2C19 and SVIL (Supervillin) gene polymorphic site detection kit and application
  • Human cytochrome P450CYP2C19 and SVIL (Supervillin) gene polymorphic site detection kit and application
  • Human cytochrome P450CYP2C19 and SVIL (Supervillin) gene polymorphic site detection kit and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0198] Example 1 Establishment of real-time fluorescent quantitative allele-specific PCR and its clinical evaluation and application

[0199] 1. Establishment of real-time fluorescence quantitative allele-specific PCR method

[0200] 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

[0249] Example 2: Detection of CYP2C19*2; CYP2C19*3; CYP2C19*17 and SVIL genotypes by sequencing

[0250] 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 SVIL 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 fluorescence quantitative PCR Taqman.

[0251] 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 populations, and...

Embodiment 3

[0296] This example provides a human cytochrome P450 CYP2C19*2: rs4244285; CYP2C19*3: rs4986893; CYP2C19*17: rs12248560; and SVIL: rs17834991 genomic single nucleotide polymorphism (SNP) real-time fluorescent quantitative PCR Taqman detection Reagent test kit.

[0297] This kit can identify and detect CYP2C19*2: rs4244285; CYP2C19*3: rs4986893; CYP2C19*17: rs12248560; and SVIL: rs17834991 SNP sites 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 flu...

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Abstract

The invention provides a cerebral apoplexy pharmacy and genetic correlation detection method. According to the detection method, primers and probes are synthesized through four polymorphic sites of two genes of cytochrome P450CYP2C19*2: rs4244285, CYP2C19*3: rs4986893, CYP2C19*17: rs12248560, and SVIL: rs17834991, real-time fluorescence quantification allele probe detection is implemented, and whether a target belongs to pharmacy susceptible populations or not can be judged. By adopting the method, clinical pharmacy schemes can be instructed and adjusted, bases can be made for clinical individual treatment, and adverse reactions of medicines are prevented. The method is capable of simultaneously detecting four polymorphic sites and has the advantages of being simple and convenient, accurate, rapid, high in flux and the like.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a detection kit for human cytochrome P450 CYP2C19 and SVIL 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 circulatio...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/12C07K14/47
CPCC12Q1/6858C12Q1/6883C07K14/47C12Q2600/16C12Q2600/156C12Q2600/106C12Q2537/143C12Q2563/107
Inventor 龙凌云田鸣
Owner 龙凌云
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