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Kit and method for detecting gene polymorphism related to warfarin personalized medication by pyro sequencing method

A technology of pyrosequencing and gene polymorphism, which is applied in the field of molecular biology, can solve problems such as cross-contamination and insufficient accuracy, and achieve the effect of small sample size and easy operation

Inactive Publication Date: 2012-09-19
周宏灏
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Problems solved by technology

[0008] Although there are currently many methods for detecting warfarin dosage-related gene polymorphisms, such as polymerase chain reaction (PCR), restriction enzyme fragment length polymorphism (RFLP), real-time fluorescence quantitative Polymerase chain reaction, nucleic acid sequence amplification experiment method, gene chip, etc. These methods are relatively fast and sensitive, but the accuracy is not enough, and the problem of cross-contamination is relatively serious

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  • Kit and method for detecting gene polymorphism related to warfarin personalized medication by pyro sequencing method
  • Kit and method for detecting gene polymorphism related to warfarin personalized medication by pyro sequencing method
  • Kit and method for detecting gene polymorphism related to warfarin personalized medication by pyro sequencing method

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Embodiment 1

[0039] VKORC1 -1639G>A (rs9923231) upstream primer: 5′-AGG GTT CAA GTG GTT CTC GT-3′ (SEQ ID NO.3);

[0040] CYP2C9*3 1075A>C (rs1057910) upstream primer: 5′- AGC CAC ATG CCC TAC ACA G -3′ (SEQ ID NO.4);

[0041] VKORC1 -1639G>A (rs9923231) downstream primer: 5′-GTC AAG CAA GAG AAG ACC TG-3′ (SEQ ID NO.5);

[0042] CYP2C9*3 1075A>C (rs1057910) downstream primer: 5′- CCC GGT GAT GGT AGA GGT TTA -3′ (SEQ ID NO.6);

[0043] VKORC1 -1639G>A (rs9923231) sequencing primer: 5′- GTG AGC CAC CGC A -3′ (SEQ ID NO.7);

[0044] CYP2C9*3 1075A>C (rs1057910) sequencing primer: 5′- CAC GAG GTC CAG AGA TA -3′ (SEQ ID NO.8);

[0045] 1. DNA extraction

[0046] 1.1 The preparation and inspection of reagent materials before the experiment are as follows:

[0047] (1) Check the shelf life of the kit and ensure that ethanol has been added to Wash Buffer 1 and 2, and tick the corresponding marks on the bottle; (2) Isopropanol (if not available, absolute ethanol can be used instead) and 75...

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Abstract

The invention discloses a kit and a method for detecting the gene polymorphism related to warfarin personalized medication by a pyro sequencing method. The kit is used for typing genes related to the warfarin medication, and the single nucleotide polymorphism of VKORC1-1639 G>A (rs9923231) and CYP2C9*31075A>C(rs1057910) is involved; and the kit comprises primers shown as SEQ ID NO.3-8. By the kit, VKORC1-1639G>A and CYP2C9*3 1075A>C can be detected accurately and quickly in high flux, so that the safe, reasonable and effective personalized administration of the warfarin medication is realized.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to a kit and a method for detecting gene polymorphism of warfarin individualized medication by pyrosequencing. Background technique [0002] Warfarin is a commonly used coumarin oral anticoagulant for the prevention and treatment of thromboembolic diseases (such as myocardial infarction, ischemic attack and thromboembolic phlebitis, etc.), as well as artificial heart valve replacement and artificial blood vessels Anticoagulant therapy after transplantation. At present, prothrombin time (PT) and international normalized ratio (INR) are often used as anticoagulation monitoring indicators in clinical practice. However, the effective therapeutic window of warfarin is very narrow, and individual differences are large (the required dose of different patients may vary by 20 times), and the initial dose is difficult to control. If the dose is insufficient, thromboembolism canno...

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 周宏灏
Owner 周宏灏
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