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Method and kit for detecting CYP2C19*2 polymorphic site genotype

A technology of CYP2C19 and polymorphic sites, which is applied in biochemical equipment and methods, microbial measurement/testing, DNA/RNA fragments, etc., can solve problems such as genotype misjudgment, and achieve the effect of improving accuracy

Inactive Publication Date: 2016-12-14
GUANGDONG PHARMA UNIV
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  • Summary
  • Abstract
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Problems solved by technology

[0016] Table 1 Misjudgment of genotypes that may occur in the experimental design without internal control restriction restriction sites

Method used

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  • Method and kit for detecting CYP2C19*2 polymorphic site genotype
  • Method and kit for detecting CYP2C19*2 polymorphic site genotype
  • Method and kit for detecting CYP2C19*2 polymorphic site genotype

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] In this example, a series of primer pairs for detecting the genotype of the CYP2C19*2 polymorphic site were designed for AvaI restriction enzyme and isozyme, and a detection method for the genotype of the CYP2C19*2 polymorphic site was established. details as follows:

[0065] 1. Genomic DNA extraction

[0066] Using oral swab genomic DNA extraction kit or blood genomic DNA extraction kit, extract genomic DNA according to the operation steps in the manual. The DNA source is not limited thereto, and any somatic cell-derived DNA is acceptable.

[0067] 2. Primer design

[0068] Get the 1000bp sequence before and after the CYP2C19*2 polymorphic site, analyze its homology with other genes of the CYP2C subfamily, namely CYP2C9, CYP2C18, and CYP2C8, and use Primer Premier 5.0 software to design specific typing for this site primers. Wherein, the upstream primer is located within 1000 bp of the 5' end of the polymorphic site, and the downstream primer is located at the 3' ...

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Abstract

The invention discloses a method and kit for detecting the CYP2C19*2 polymorphic site genotype. During the process of detecting CYP2C19*2 polymorphic site genotype, a specific internal quality control mechanism is introduced, namely, an internal control enzyme cutting site is introduced into a PCR product; according to the restriction map, the incomplete enzyme cutting can be identified; moreover, under the situation that the enzyme cutting is incomplete and part of enzyme cutting strips is obscure, the genotype can still be judged accurately, and the accuracy of detection results is greatly improved.

Description

technical field [0001] The invention belongs to the field of molecular detection, and in particular relates to a method and a kit for detecting the genotype of a CYP2C19*2 polymorphic site. Background technique [0002] CYP2C19 is an important member of the cytochrome P450 (Cytochrome P450, CYP) enzyme system and an important drug metabolizing enzyme in the body. It was first isolated from human liver by Wrighton et al. in 1993. The full length of its cDNA is 1940bp, and the coding region is 1473bp , encoding 490 amino acid residues. CYP2C19 participates in a wide range of drug metabolism, including proton pump inhibitors (omeprazole, lansoprazole, pantoprazole), antidepressants (fluoxetine, citalopram, escitalopram, albino Mitriptyline, clomipramine, imipramine, moclobemide, trimipramine, etizolam), antiepileptic drugs (valium, clobazam, phenytoin, phenobarbital, valproic acid ), platelet aggregation inhibitors (clopidogrel), antimalarials (proguanil), antifungals (vorico...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6876C12Q1/686C12Q2600/106C12Q2521/301C12Q2565/125
Inventor 何震宇陈珂蕊谢小妃孙丹晨
Owner GUANGDONG PHARMA UNIV
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