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Primer group for detecting polymorphism of human cytochrome P450 related gene, kit and detection method

A gene polymorphism and cytochrome technology, applied in the field of primer sets of gene polymorphism sites, can solve the problems of nucleic acid loss, increase cross-contamination, use of toxic reagents, etc., to reduce pain, reduce blood consumption, and reduce economy. effect of spending

Inactive Publication Date: 2018-11-20
XIAMEN WIZ BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

During fluorescent quantitative PCR amplification, nucleic acid extraction and purification must be performed on blood or saliva in advance, resulting in a large loss of nucleic acid, increased cross-contamination, and the use of toxic reagents, etc.

Method used

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  • Primer group for detecting polymorphism of human cytochrome P450 related gene, kit and detection method
  • Primer group for detecting polymorphism of human cytochrome P450 related gene, kit and detection method
  • Primer group for detecting polymorphism of human cytochrome P450 related gene, kit and detection method

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Experimental program
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Effect test

Embodiment 1

[0056] Embodiment 1: the preparation of kit

[0057] 1. Design and synthesis of primers and probes

[0058] For the CYP2D6*10, CYP2C9*3, CYP2C19*2, CYP2C19*3, CYP3A4*18 and CYP3A5*3 gene loci in the human genome (for the sequence, see the human whole genome sequence published in the NCBI database), use PrimerPremier 3.0 and BeaconDesigner software, Design specific primers and probes. Specific primers and probe sequences are shown in Table 1:

[0059] Table 1:

[0060]

[0061]

[0062] 2. Selection of reference substance

[0063] The wild-type control is the wild-type plasmid mixture of CYP2D6*10, CYP2C9*3, CYP2C19*2, CYP2C19*3, CYP3A4*18 and CYP3A5*3; the negative control is DNA solution (TE buffer).

[0064] 3. Composition of PCR reaction solution

[0065] The PCR reaction solution includes the above-mentioned specific primers and probes, and is divided into reaction tube A and reaction tube B, wherein:

[0066] Reaction tube A contains CYP3A4*18 forward primer 9...

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Abstract

The invention discloses a primer group for detecting polymorphism of a human cytochrome P450 related gene. The primer group comprises specific primers and probes aiming at gene sites of CYP2C19*2, CYP2C19*3, CYP3A4*18, CYP2C9*3, CYP2D6*10 and CYP3A5*3. The invention further relates to a kit for detecting polymorphism sites of the human cytochrome P450 related gene. The kit comprises PCR (Polymerase Chain Reaction) reaction liquid freeze-dried powder (PCR buffer, dNTPs, MgCl2, UNG enzyme, Taq enzyme and trehalose) with the primers and the probes. The invention further relates to a method for detecting polymorphism sites of the human cytochrome P450 related gene. By adopting the method, a DNA (Deoxyribonucleic Acid) extraction procedure is avoided, micro whole-blood amplification is directlyimplemented, a small amount of blood is used, and patients can be relieved from pain; the time and the labor can be saved, and in addition, economic expense can be greatly reduced.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a primer set, a kit and a detection method for detecting gene polymorphic sites related to human cytochrome P450. Background technique [0002] After the drug enters the human body, it will go through four processes: absorption, distribution, metabolism, and excretion. The liver is the most important organ for metabolizing drugs in the human body, and the main metabolic enzymes are the cytochrome P450 superfamily (CYP), which catalyze the metabolism of various endogenous and exogenous substances (including most clinical drugs), and participate in half of the human body. The above drug metabolism process. A DNA sequence polymorphism caused by a single nucleotide variation is called a single nucleotide polymorphism. Single nucleotide polymorphisms in P450 metabolic enzymes are key contributors to differences in human metabolism and response. The genes of P450 enzymes mainly include ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/106C12Q2600/156
Inventor 杨璐平林志铿王松林雷洋
Owner XIAMEN WIZ BIOTECH CO LTD
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