Detection system for detecting CYP2D6*10 gene mutation and kit of detection system

A kit and system technology, applied in the field of detection products for detecting CYP2D6*10 gene mutations, can solve problems affecting the accuracy of detection results, difficulty in large-scale promotion and application, and cumbersome operation processes, etc., to achieve absolutely accurate quantitative analysis and improve The effect of sensitivity, good accuracy and reproducibility

Pending Publication Date: 2018-11-13
上海赛安生物医药科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, technologies such as next-generation sequencing, FLP, allele-specific oligonucleotide hybridization, allele-specific PCR, and gene chips are mainly used, but the operation process is cumbersome, and reaction tubes need to be replaced, which is prone to pollution. post-processing, affecting the accuracy of the test results
Or the instruments and equipment are expensive, and it is difficult to promote and apply them on a large scale

Method used

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  • Detection system for detecting CYP2D6*10 gene mutation and kit of detection system
  • Detection system for detecting CYP2D6*10 gene mutation and kit of detection system
  • Detection system for detecting CYP2D6*10 gene mutation and kit of detection system

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Embodiment 1

[0040] Example 1 Application of Detection Kit for CYP2D6*10 Gene Mutation

[0041] 1. The composition of the kit.

[0042] The CYP2D6*10 gene mutation detection kit of this embodiment includes digital PCR premix, primer-probe mixture, positive quality control and negative quality control. The components of the kit are shown in Table 1.

[0043] Table 1 Kit composition list

[0044] Element

pack

Reagent company

Digital PCR Master Mix

1 tube

Life technologies

Primer Probe Mix

1 tube

——

positive control

1 tube

Shanghai Bailige Biotechnology Co., Ltd.

negative control

1 tube

Shanghai Bailige Biotechnology Co., Ltd.

[0045] The description of the kit components in the above Table 1 is as follows:

[0046] 1) The main components of the digital PCR master mix include DNA polymerase, ultrapure water, dNTPs mixture, droplet stabilizer and buffer (provided by Life technologies, product number: 1508099),...

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Abstract

The invention relates to a detection system for detecting CYP2D6*10 gene mutation and a kit of the detection system. The detection system comprises digital PCR (polymerase chain reaction) premixed solution and primer probe mixed solution, the primer probe mixed solution comprises an upstream primer, a downstream primer and a peptide nucleic acid probe which are used for detecting CYP2D6*10 sites of CYP2D6 genes, a nucleotide sequence of the upstream primer is as shown in SEQ ID No.1, a nucleotide sequence of the downstream primer is as shown in SEQ ID No.2, a nucleotide sequence of the peptidenucleic acid probe and CYP2D6*10 mutant gene sites are completely complemented, the nucleotide sequence of the peptide nucleic acid probe is as shown in SEQ ID No.3, and the 5' end of the peptide nucleic acid probe is connected with a strong chelation group formed by four amino acids. According to the detection system for detecting CYP2D6*10 gene mutation and the kit of the detection system, sensitivity and specificity are high, needed samples are less, and quantitative PCR detection can be performed rapidly, conveniently and accurately.

Description

technical field [0001] The invention relates to a detection product for detecting CYP2D6*10 gene mutation, which belongs to the field of biotechnology. Background technique [0002] Isoquine 4'-hydroxylase (CYP2D6) is an important member of the second subfamily of the cytochrome P450 superfamily (cytochrome P450s, CYP). CYP2D6*10 genotype polymorphism is closely related to the survival rate of breast cancer patients treated with tamoxifen (TAM). The activity of CYP2D6 in the population presents a four-state distribution phenomenon of intensive metabolizers (EM), intermediate metabolizers (IM), poor metabolizers (PM) and ultra metabolizers (UM). At present, more than 100 mutant genes of CYP2D6 have been found, and its mutation type determines the metabolic activity of the enzyme. CYP2D6 gene polymorphism determines the difference in drug metabolism intensity, and affects the selection of clinical drug varieties and doses. In addition to participating in the metabolism of s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6858C12Q1/6851
CPCC12Q1/6851C12Q1/6858C12Q2537/143C12Q2563/159C12Q2525/107
Inventor 王明赵国玲洪专
Owner 上海赛安生物医药科技股份有限公司
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