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A method for detection of complete gene deletion of cyp2a6 based on fluorescent quantitative PCR

A fluorescent quantitative, whole-gene technology, applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of difficult combination of primers and probes, save experimental time and consumables, ensure accuracy, and improve sensitivity Effect

Active Publication Date: 2015-12-02
SHANXI LIFEGEN
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Problems solved by technology

[0007] However, this technology has not been applied to the detection of CYP2A6 gene deletion; the fundamental reason is that the CYP2A6 gene has a very high sequence homology (more than 90%) with the CYP2A7 gene and CYP2A13 gene. It is very difficult to combine primers and probes to amplify CYP2A6 gene with high specificity in PCR reaction

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  • A method for detection of complete gene deletion of cyp2a6 based on fluorescent quantitative PCR
  • A method for detection of complete gene deletion of cyp2a6 based on fluorescent quantitative PCR
  • A method for detection of complete gene deletion of cyp2a6 based on fluorescent quantitative PCR

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Embodiment Construction

[0042]The present invention provides a method for quickly and accurately detecting CYP2A6 gene deletion based on fluorescent quantitative PCR, that is, by designing highly specific primers for CYP2A6 gene and ALB (as an internal reference gene) respectively, and using fluorescent quantitative PCR technology to quickly and accurately detect CYP2A6 A method for whole gene deletion.

[0043] see figure 1 As shown, the specific primers and probe combinations of the CYP2A6 gene designed in the present invention. Based on the slight difference in the sequences of CYP2A6 gene and CYP2A7 and CYP2A13 genes, and based on the principle that the 3' end sequence of the primer determines its extension efficiency, we designed a set of primer-probe combinations that can amplify the CYP2A6 gene with high specificity. Using this combination for real-time quantitative PCR can not only efficiently amplify the CYP2A6 gene, but also completely avoid the interference of the CYP2A7 and CYP2A13 genes...

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Abstract

The invention provides a method for detecting CYP2A6 whole-gene deletion based on a fluorescent quantitative PCR. The method is used for achieving the aim of non-disease diagnosis. Highly-specific primers for CYP2A6 genes and ALB genes are respectively designed, and CYP2A6 whole-gene deletion is detected fast and accurately based on the fluorescent quantitative PCR technology. Primer probe combinations for CPY2A6 comprise Fp:5'-TCCCACCCTACTCCCTCTCTC-3', Rp:5'-GGAGGTGAACGCGGGA-3' and Probe:5'-FAM-TGGAGACAGGCCAGGGGGCG-BHQ2-3'.

Description

technical field [0001] The invention relates to a method for detecting CYP2A6 whole gene deletion. Background technique [0002] The CYP2A6 gene is a member of the human cytochrome P450 gene superfamily, and the protein encoded by it is an important metabolic enzyme involved in the metabolism of various drugs and toxic compounds in the human body. The CYP2A6 gene shows a wide range of polymorphisms in the population, and the frequency of CYP2A6 gene deletion in the Chinese population is 5%-15%. [0003] Tegafur and compounds with tegafur as the main active ingredient are currently the most widely used anti-pyrimidine drugs in clinical practice, and have good therapeutic effects on digestive tract tumors and other solid tumors. [0004] A large number of studies have shown that the CYP2A6 gene deletion polymorphism is closely related to the efficacy of drugs such as S-1 and UFT with tegafur as the active ingredient. Patients without the polymorphism had higher drug response...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6851C12Q1/6858C12Q2531/113C12Q2563/107C12Q2561/113
Inventor 戴鹏高寻晓洁陈超邹晖
Owner SHANXI LIFEGEN
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