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Method and kit for detecting CYP2C19 genotyping based on real-time fluorescent quantitative PCR

A CYP2C19, real-time fluorescence quantitative technology, applied in the field of bioengineering, can solve the problems of cumbersome operation steps and high cost, and achieve the effect of simple operation, low cost and high accuracy

Inactive Publication Date: 2020-09-01
武汉翌圣生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the cost of this method is high and the operation steps are cumbersome

Method used

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  • Method and kit for detecting CYP2C19 genotyping based on real-time fluorescent quantitative PCR
  • Method and kit for detecting CYP2C19 genotyping based on real-time fluorescent quantitative PCR
  • Method and kit for detecting CYP2C19 genotyping based on real-time fluorescent quantitative PCR

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Embodiment 1

[0028] The design of embodiment 1 primer pair and probe

[0029] 1. Search for CYP2C19 SNP site sequence

[0030] In this example, the SNP sites of CYP2C19 were searched on the dbSNP database, and the *2 and *3 sites were selected for primer and probe design. The URL of the dbSNP database in this embodiment is: http: / / www.bioinfo.org.cn / relative / dbSNP%20Home%20Page.htm.

[0031] 2. Design primer pairs and probes according to the design principles of primers and probes, as shown in Table 1.

[0032] Table 1 Primer pairs and probes

[0033]

[0034] Among them, the amplification length of CYP2C19*2 is 244bp; the amplification length of CYP2C19*3 is 235bp.

[0035] 3. qPCR amplification system

[0036] The probes synthesized at each of the above SNP sites were mixed into 10 uM TaqMan Probe mix, and each site was configured into a 20 μL system according to the formula in Table 2 for reaction.

[0037] Table 2 qPCR amplification system

[0038]

[0039]

[0040] 4. q...

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Abstract

The invention discloses a method for detecting CYP2C19 genotyping based on real-time fluorescent quantitative PCR (Polymerase Chain Reaction) for non-disease treatment and diagnosis purposes, comprising the following steps of: 1, performing fluorescent quantitative PCR amplification by taking DNA (Deoxyribonucleic Acid) of a sample to be detected as a template to obtain a PCR amplification product; 2, detecting a fluorescence signal of an amplification product; 3, judging whether a sample contains the CYP2C19 * 2 genotype or not, according to a detection result, wherein a reaction system for PCR amplification contains a specific primer pair for amplifying the CYP2C19 * 2 gene and a fluorescent probe for detecting the CYP2C19 * 2 gene. The invention also discloses a primer pair and a fluorescent probe for detecting the CYP2C19 genotyping. The method provided by the invention has high specificity and high detection result accuracy.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to a method for detecting CYP2C19 genotyping based on real-time fluorescence quantitative PCR and a kit thereof. Background technique [0002] CYP2C19 is an important member of the second subfamily of CYP450 enzymes. Located on chromosome 10, the protein composed of 490 amino acids is expressed in many tissues of organisms, mainly in liver tissue. Currently, at least 10% of drugs act on it in clinical application. CYP2C19 has many SNP sites. At present, among the 25 mutant alleles of CYP2C19 that have been found, at least 10 mutations cause changes in enzyme activity. The slow metabolizers are mainly CYP2C19*2 and CYP2C19*3, and the fast metabolizers The type is mainly CYP2C19*17. [0003] Asians and Pacific Islanders have a higher probability of non-functional mutations for CYP2C19, which may affect the treatment of related drugs, such as the antiplatelet drug ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12Q1/686C12N15/11
CPCC12Q1/6883C12Q1/686C12Q2600/106C12Q2600/156C12Q2561/113C12Q2563/107C12Q2545/114C12Q2561/101
Inventor 周其好严广号孙永祥宋东亮李剑辉
Owner 武汉翌圣生物科技有限公司
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