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SYBR Green I detection primer of CYP2D6*10 gene, kit and detection method thereof

A detection kit and detection primer technology, applied in the medical and biological fields, can solve the problems of high false positive rate, low sensitivity, long time-consuming, etc., and achieve the effects of high sensitivity, simple operation, and overcoming easy pollution.

Pending Publication Date: 2020-02-21
爱尔生基因医学科技有限公司
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Problems solved by technology

However, the Taqman fluorescent probe method has low sensitivity, high false positive rate, long time-consuming, and low throughput

Method used

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  • SYBR Green I detection primer of CYP2D6*10 gene, kit and detection method thereof

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Embodiment 1

[0027] 1. Synthetic detection primers and positive control sequences

[0028] The selected primers of the present invention are designed for the CYP2D6*10 gene polymorphism. The selected primers are from a set of primer combinations artificially added with mismatched mutation sites at the 3' end of a sequence and its vicinity. By screening primers that can distinguish alleles of a single site, the height of PCR amplification is ensured. specificity.

[0029] After experimental verification, the specific detection primers for CYP2D6*10 gene polymorphism detection were finally determined, and the nucleotide sequences are as follows:

[0030] CYP2D6*10 gene wild-type upstream primer (F1), mutant upstream primer (F2) and common downstream primer (R1)

[0031] F1 (SEQ ID NO: 1): 5`-tgtgcggactcctaaatccgc-3`

[0032] F2 (SEQ ID NO:2): 5`-gtggtctccgtgagtctccg-3`

[0033] R1 (SEQ ID NO:3): 5`-gtctcggactcataaatcctg-3`

[0034] The wild-type positive control of the CYP2D6*10 gene co...

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Abstract

The invention provides an SYBR Green I detection primer of a CYP2D6 * 10 gene, a kit and a detection method. Wherein the detection primer comprises a CYP2D6*10 wild type upstream primer, a CYP2D6*10 mutant type upstream primer, a CYP2D6*10 wild type universal downstream primer and a CYP2D6*10 mutant type universal downstream primer; the nucleotide sequence of the CYP2D6*10 wild type upstream primer is shown as SEQ ID NO: 1, the nucleotide sequence of the CYP2D6*10 mutant type upstream primer is shown as SEQ ID NO: 2, and the nucleotide sequence of the universal downstream primer is shown as SEQ ID NO: 3. According to the application, the detection primers and the kit containing the primers are used; a fluorescence signal value in the PCR process is directly explored, so that the detectionresult is obtained, PCR post-treatment or electrophoresis detection is not needed, the technical problems of easy pollution and false positive of the conventional PCR technology are solved, the problem of non-specific amplification can be effectively avoided, and the method is suitable for detection of large-batch samples. The method has the advantages of high sensitivity and strong specificity, and can guide patients with various diseases to realize individualized medication according to individual genome information.

Description

technical field [0001] The invention relates to the fields of medicine and biotechnology, in particular to a SYBR Green I detection primer, kit and detection method for CYP2D6*10 gene. Background technique [0002] The cytochrome P450 (CYP) family is an important enzyme system in the phase I metabolism of drugs, and plays an important role in the metabolism of endogenous and exogenous substances in organisms. The CYP2D6 gene is one of the important members of the CYP enzyme family. Although it only accounts for 2-9% of the total liver enzymes, it participates in the metabolism of 20-30% of drugs, including antidepressants, antiarrhythmics, antipsychotics, and sedatives. Pain medicine etc. The activity of CYP2D6 in different individuals can vary up to 1000 times. The most common polymorphism in the Chinese population is CYP2D6*10. About 51.6% of Chinese people carry this genetic variation. Unstable, thus affecting drug metabolism. [0003] By detecting the patient's CYP2D6...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/6858C12N15/11
CPCC12Q1/6888C12Q1/6858C12Q2600/106C12Q2600/156C12Q2531/113C12Q2563/107C12Q2545/113
Inventor 谢少华
Owner 爱尔生基因医学科技有限公司
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