Fluorescence quantitative detecting method for CYP2C19 genotyping

A CYP2C19, fluorescence quantitative detection technology, applied in the field of gene detection and drug metabolism analysis, can solve the problems of time-consuming operation and low throughput

Inactive Publication Date: 2009-09-09
樊世斌 +1
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Problems solved by technology

[0020] The object of the present invention is to provide a kind of CYP2C19 genotyping fluorescent quantitative detection method and diagnostic kit; It combines the characteristics of ASA detection SNP and single base mutation with TaqMan fluorescent quantitative detection characteristics, solves the problem of conventional gel electrophoresis method judgment The problems of time-consuming operation and low throughput faced by ASA results

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  • Fluorescence quantitative detecting method for CYP2C19 genotyping
  • Fluorescence quantitative detecting method for CYP2C19 genotyping

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[0065] CYP2C19 enzyme activity affects the metabolism of various drugs such as omeprazole. In the Chinese population, the low metabolic phenotype is almost entirely determined by the M1 and M2 mutations. Detection of M1 and M2 mutations can determine enzyme activity and predict the in vivo metabolism and side effects of related drugs. So far, ASA technology is mostly used in typing methods, and gel electrophoresis is used to judge ASA results. The operation is time-consuming and the throughput is low, which is difficult to meet the needs of clinical routine testing. Therefore, the present invention proposes a new method of replacing gel electrophoresis with TaqMaq fluorescence quantification, without gel electrophoresis, and real-time quantitative analysis of CYP2C19 genotype.

[0066] The integration of the two technologies refers to: in the quantitative PCR reaction system, different ASA primer pairs, TaqMan-MGB probes and corresponding PCR reagents are used to perform paral...

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Abstract

The invention relates to a fluorescent quantitative detecting method for CYP2C19 genotyping fluorescent and a diagnostic kit. At present, CYP2C19 genotyping techniques at home and abroad are mainly RFLP technique and ALA technique which determine genotypes according to the size of a DNA fragment, with the disadvantages of time-consuming operation and low flux. The invention provides a novel point mutating or SNP detecting method which utilizes the ASA combination primer sequence designed for the polymorphic loci of the CYP2C19 gene exons 4 and 5, the referential ASA primer combination or the degenerated primer sequence for quality control, specific TaqMan-MGB probe sequences for amplified products, an ASA amplifying reaction method, the amplifying reaction result of real-time fluorescent quantitative detection, the fast analysis of mutation locus type and genotyping. The quantitative detecting method has the advantages of time saving compared with a conventional ASA method, no need for electrophoresis detection, fastness, accuracy, and the like, and can be used for detecting other drug-metabolizing enzymes, or more extensive genetic variation or mutation.

Description

technical field [0001] The invention belongs to the technical field of gene detection and drug metabolism analysis, and specifically relates to a CYP2C19 genotype fluorescence quantitative detection method. Background technique [0002] 1. CYP2C19 function and its gene polymorphism [0003] Cytochrome P450 is an isozyme encoded by a group of superfamily genes. In humans, it is mainly present in hepatocyte microsomes. Among them, CYP2C19 catalyzes the hydroxylation metabolism of S-Mephenytoin, so it is also called S-Mephenytoin Hydroxylase, which was isolated from human liver by WrJighton et al. in 1993 and participates in the metabolism of various drugs. [0004] CYP2C19 participates in a variety of clinical drugs through hydroxylation, oxidation, and cyclization reactions (such as mephenytoin, omeprazole, diazepam, propranolol, propranolol, imipramine, clomipramine, proguanil, etc.) metabolism. There are genetic differences in the hydroxylation metabolism of mephenytoin...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 樊世斌白玉杰
Owner 樊世斌
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