Kit and method for detecting polymorphism of CYP2D6 gene

A gene polymorphism and kit technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of long time, long time, and complicated steps for CYP2D6 gene polymorphism, so as to save detection time. , high accuracy, and the effect of reducing pollution

Active Publication Date: 2015-05-13
刘辉
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  • Claims
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AI Technical Summary

Problems solved by technology

[0005] In view of the problems in the prior art that the detection of CYP2D6 gene polymorphism takes a long time, the steps are cumbersome, and multiple PCRs are required, the purpose of the present invention is to provide a kit for detecting CYP2D6 gene polymorphism
[0007] In order to solve the problems in the prior art that it takes a long time to detect CYP2D6 gene polymorphism, the steps are cumbersome, and multiple PCRs are required, the present invention adopts the following technical solutions:

Method used

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  • Kit and method for detecting polymorphism of CYP2D6 gene
  • Kit and method for detecting polymorphism of CYP2D6 gene
  • Kit and method for detecting polymorphism of CYP2D6 gene

Examples

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Embodiment 1

[0050] Embodiment 1. Preparation of kit

[0051] 1. Primer design for internal reference gene ACTB and CYP2D6 gene

[0052] According to the gene sequence, the internal reference gene ACTB sequence and the CYP2D6 gene sequence are derived from the National Center for Biotechnology Information Nucleic Acid Database (NCBI), wherein the internal reference gene ACTB ID is 60, and the reference sequence number is NG_007992.1, also refer to SEQ in the sequence list of the present invention ID NO: 15; CYP2D6 gene ID is 1565, and the reference sequence number is NG_008376.2. Primer5.0 primer design software was used to design specific primers for the internal reference gene ACTB and CYP2D6 gene, including: exon 1 to exon 9 of CYP2D6 gene The upstream and downstream primers of the full-length amplification specific primers of exons; the upstream and downstream primers of the first exon sequencing primers of CYP2D6 gene; the upstream and downstream primers of the second exon sequencing ...

Embodiment 2

[0073] Example 2. Detection of CYP2D6 gene polymorphism with the kit prepared in Example 1

[0074] Take the results of random detection of peripheral blood samples from 30 subjects as an example.

[0075] The detection process of using the kit of the present invention to detect CYP2D6 gene polymorphism in the population is as follows: firstly, obtain the peripheral blood sample of the clinical subject, and quickly extract the genomic DNA; secondly, first prepare the PCR reaction solution of the internal reference gene ACTB, and then add the concentration Both 2ng / μl internal reference gene ACTB sequence and 2μl internal positive control sequence were used for PCR amplification. After PCR, electrophoresis was performed in agarose DNA gel with a concentration of 2% to check the PCR amplification situation. If there are two bands of about 390bp and 800bp in the PCR amplification product of the internal reference gene ACTB, it indicates that the entire detection process is valid....

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Abstract

The invention relates to a kit and method for detecting the polymorphism of a CYP2D6 gene and belongs to the technical field of gene sequencing. The kit comprises primers for detection, wherein the primers for the detection include full-length amplification specific primers for a first exon to a ninth exon of the CYP2D6 gene, and at least one pair in forward and reverse sequencing primers for the first exon of the CYP2D6 gene, forward and reverse sequencing primers for the second exon of the CYP2D6 gene, forward and reverse sequencing primers for the third exon and the fourth exon of the CYP2D6 gene, forward and reverse sequencing primers of the fifth exon to the seventh exon of the CYP2D6 gene, and forward and reverse sequencing primers of the eighth exon and the ninth exon of the CYP2D6 gene. By the adoption of the kit and the method, the detection time is obviously shortened and the workload is remarkably reduced. The kit provides a brand-new quick and simple way for prediction of the dosage of a medicine which is metabolized by CYP2D6.

Description

technical field [0001] The invention relates to the technical field of gene sequencing, in particular to a kit and method for detecting CYP2D6 gene polymorphism. Background technique [0002] Cytochrome P450 (CYP450) is a kind of gene family composed of heme-thiolate protein, which is mainly distributed on the endoplasmic reticulum and mitochondrial inner membrane. In addition to participating in the synthesis of sterol hormones in organisms, it can also participate in the Metabolism of endogenous and exogenous substances. The structure, function and role of cytochrome P450 in drug metabolism have been extensively studied at home and abroad. In the important cytochrome P450 family in pharmacogenetics, the enzyme encoded by the CYP2D6 gene is one of its members, and the gene is located at 22q13.1. Studies have shown that it is involved in more than 25% of commonly used drugs such as isoquine (adrenergic blocking drug), spatine and propafenone (antiarrhythmic drug) and amitr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 刘辉
Owner 刘辉
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