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CYP2D6*10 genetic polymorphism detection kit capable of distinguishing CYP2D7P and CYP2D8P

A CYP2D6-F, CYP2D6-R technology, applied in the field of molecular medicine, can solve problems such as interference of genotyping results, and achieve the effects of pure amplification products without interference, rapid detection and high sensitivity

Active Publication Date: 2021-04-23
XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the existence of pseudogenes, it is difficult for CYP2D6 to be amplified by ordinary primers like other genes without pseudogenes. Often, in addition to CYP2D6, the pseudogenes of the amplified products are also expanded together, making the genotyping results Severe interference, which is also an interference factor that many studies have not focused on

Method used

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  • CYP2D6*10 genetic polymorphism detection kit capable of distinguishing CYP2D7P and CYP2D8P
  • CYP2D6*10 genetic polymorphism detection kit capable of distinguishing CYP2D7P and CYP2D8P
  • CYP2D6*10 genetic polymorphism detection kit capable of distinguishing CYP2D7P and CYP2D8P

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Embodiment 1

[0028] 1. Materials

[0029] 1. Instrument

[0030] Real-time fluorescent PCR instrument, pipette, centrifuge, small vortex instrument, UV spectrophotometer.

[0031] 2. Primers and probes

[0032] In this example, a pair of high-efficiency specific primers are designed for the detection of target genes, and a pair of labeled fluorescent dye probes are designed to detect the corresponding CYP2D6 polymorphic sites.

[0033]The sequences of primers and probes are as follows:

[0034] Primers:

[0035] CYP2D6-F:5'-GCGCTCGGTGTGCT-3' (SEQ ID NO.1)

[0036] CYP2D6-R:5'-CTGTGGTTTCACCCCACCAT-3' (SEQ ID NO.2);

[0037] Probe:

[0038] CYP2D6-Pw:FAM-5'-CTGCACGCTACCCACC-3'-BHQ1 (SEQ ID NO.3)

[0039] CYP2D6-Pm:HEX-5'-CTGCACGCTACTCACC-3'-BHQ1 (SEQ ID NO. 4).

[0040] 3. Reagents

[0041] 10mmol / L Tris-HCl pH 8.0, 2.5mmol / L MgCl 2 , 2.0mmol / L dNTP solution, 1UChampagne Taq, 0.1U UNG enzyme.

[0042] 2. Method

[0043] 1. Sample selection

[0044] Whole blood samples from the p...

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Abstract

The invention discloses a CYP2D6*10 genetic polymorphism detection kit capable of distinguishing CYP2D7P and CYP2D8P, and belongs to the technical field of molecular medicine. The kit can utilize a highly specific primer pair CYP2D6 (rs1065852) to realize accurate target amplification; and through the combination of the advantages of a Taqman probe of being high in specificity, fast and simple, the genotyping of the CYP2D6 (rs1065852) can be realized.

Description

technical field [0001] The invention belongs to the technical field of molecular medicine, and specifically relates to a detection kit for distinguishing pseudogenes CYP2D7P and CYP2D8P so as to accurately amplify and type CYP2D6*10 genetic polymorphism. Background technique [0002] Cytochrome P450 2D6, also known as isoquinidine 4'-hydroxylase, is an important member of the second subfamily of CYP, an enzyme encoded by the human CYP2D6 gene, mainly expressed in the liver and the central nervous system. CYP2D6 metabolizes and eliminates about 25% of clinically used drugs through hydroxylation, demethylation, and dealkylation, and is one of the most important drug-metabolizing enzymes in the human body. [0003] Due to the different genotypes of CYP2D6 among different individuals, there are great differences in the activity and quantity of its enzyme. Metabolizer (UM) four-state distribution phenomenon. For drugs metabolized by CYP2D6, ie, CYP2D6 substrates, some individua...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/6858C12N15/11
CPCC12Q1/6888C12Q1/6858C12Q2600/156C12Q2600/106C12Q2531/113C12Q2561/101C12Q2563/107
Inventor 方逸楠庄江兴张弦杨万宝
Owner XIAMEN UNIV
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