Method and kit for detection of human CYP2C19 gene polymorphism

A technology of CYP2C19 and gene polymorphism, applied in biochemical equipment and methods, measurement/testing of microorganisms, DNA/RNA fragments, etc., can solve the problems of high price of detection instruments, difficulty in clinical application and promotion, and high detection cost, and achieve Save costs, reduce the possibility of pollution, and ensure the effect of accuracy

Inactive Publication Date: 2017-09-01
SHANGHAI BIOMED UNION BIOTECHNOLOGY CO LTD
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Problems solved by technology

[0016] In view of this, the present invention provides a new method and kit for detecting human CYP2C19 gene polymorphisms to solve the problems of high price, complicated operation, easy pollution and high detection cost of the detection equipment existing in previous detection technologies and finished product detection kits. Difficulty in promoting clinical application

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  • Method and kit for detection of human CYP2C19 gene polymorphism
  • Method and kit for detection of human CYP2C19 gene polymorphism
  • Method and kit for detection of human CYP2C19 gene polymorphism

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Embodiment 1

[0058] The specific detection steps of the kit for detecting human ApoE gene polymorphism of the present embodiment are as follows:

[0059] 1. DNA extraction.

[0060] Use the blood sample DNA extraction kit to extract samples 1, 2, and 3 (samples 1, 2, and 3 are all EDTA anticoagulated whole blood). For specific operations, refer to the DNA kit extraction product manual.

[0061] 2. Sample DNA quality testing.

[0062] After the sample DNA is obtained, the quality of the sample is controlled by measuring the concentration and the ratio of OD260 / OD280, and finally added to the sample in the reaction system. The optimal reaction result can be obtained when the ratio of OD260 / OD280 is between 1.7 and 2.0, and the concentration is 5-20ng / μl is more suitable.

[0063] 3. PCR reaction.

[0064] 1) Prepare 20μL PCR reaction system:

[0065] Calculate the number of PCR reaction tubes required for detection of c.681 site (Number of PCR reaction tubes = number of samples + positi...

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Abstract

The invention provides a novel method for detection of human CYP2C19 gene polymorphism. The method comprises the following steps: (1) an ARMS primer is designed. The 3' end of the ARMS primer is a mutation site, and any missense in first-third nucleotide positioned upstream of a mutation site is mutated; (2) after allelic ARMS primer is designed for each SNP, an artificial nucleotide sequence with no homology with human gene sequence is respectively introduced into the 5' end of each of the two specific ARMS primers; (3) according to the introduced general artificial sequence, a Taqman probe which is completely complementary and paired up with the intermediate sequence fragment of the general sequence is designed, and a specific fragment amplified by the ARMS primer is specifically identified. By the above detection method and the kit, the capability of the allelic specific primer in distinguishing different alleles can be greatly enhanced; and accuracy of detection results is guaranteed, cost is greatly saved and efficiency is raised.

Description

technical field [0001] The invention relates to a method for detecting gene polymorphism, in particular to a method and kit for detecting human CYP2C19 gene polymorphism. Background technique [0002] 1. Brief introduction of CYP2C19 gene [0003] Cytochrome P450 (Cytochome P450, CYP450) is a group of structurally and functionally related isoenzymes encoded by superfamily genes, and is a group of heme-containing enzymes. It is mainly involved in the related metabolism of drug I, and 80% of the drug metabolism is related to it. Among them, CYP2 is the largest family, with 15 subfamilies (CYP2A-2Q), and about 20% of clinically metabolized drugs are metabolized by it. CYP2C19 is one of the most important drug-metabolizing enzymes in the CYP2 family, and many endogenous substrates, And about 2% of clinical drugs are catalyzed by it. [0004] The CYP2C19 gene is located on the q24.1-q24.3 zone of human chromosome 10, and its entire sequence contains 9 exons, encoding 490 amino...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6858C12Q1/6883C12Q2600/106C12Q2600/156C12Q2531/113C12Q2545/114C12Q2535/137C12Q2561/101C12Q2563/107
Inventor 刘振吴国祥韩武梅齐善康金芬芬
Owner SHANGHAI BIOMED UNION BIOTECHNOLOGY CO LTD
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