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Detection kit and detection method for individualized medication of warfarin

A detection kit and detection method technology, applied in the fields of molecular biology and nucleic acid detection, can solve the problems of long test time, complicated operation and high cost, and achieve the effects of short detection time, low detection cost and low cost

Inactive Publication Date: 2015-10-07
BEIJING JINQI BIOLOGICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The problem to be solved by the present invention is that the existing methods for detecting CYP2C9 and VKORC1 genotypes have high cost, complex operation and long testing time

Method used

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  • Detection kit and detection method for individualized medication of warfarin
  • Detection kit and detection method for individualized medication of warfarin
  • Detection kit and detection method for individualized medication of warfarin

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Embodiment Construction

[0064] In order to better understand the present invention, the present invention will be further described below in conjunction with specific embodiments and accompanying drawings.

[0065] Warfarin individualized drug detection kit and method thereof, the kit includes PCR buffer, Taq enzyme, dNTP, DNA ladder Marker. Wherein, the kit provided by the invention also includes the following primers:

[0066] (1) The first round of PCR at the CYP2C9*2 locus

[0067] Upstream primer: 5'-CTGTCTACTTTTCCTAGCTCTCAAAGG-3' (SEQ ID No.1, hereinafter referred to as 2C9-F),

[0068] Downstream primer: 5′-GATAGGGCCATTCCCACCATGTTG-3′ (SEQ ID No.2, hereinafter referred to as 2C9-R);

[0069] (2) The second round of PCR at the CYP2C9*2 site

[0070] Upstream primer: 5'-TTTTCCCACTGGCTGAAAGAGCTAA-3' (SEQ ID No.3, hereinafter referred to as P2),

[0071] Downstream primer: 5'-TGGGAAAAACACTGCTCTTTAACTC-3' (SEQ ID No.4, hereinafter referred to as Q2),

[0072] Mutant-specific primer: 5'-AAGAGGA...

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Abstract

The invention provides a detection kit and detection method for individualized medication of warfarin, belonging to the technical fields of molecular biology and nucleic acid detection. According to the detection method provided by the invention, mutant types of CYP2C9 and VKORC1 are detected by using a bidirectional allele-specific polymerase chain reaction (Bi-ASPCR) method; two cycles of PCR are carried out on site CYP2C9*2 and site VKORC1-1639; one cycle of PCR is carried out on site CYP2C9*3; and obtained PCR products are subjected to electrophoresis and electrophoresis results are compared with standards. According to the invention, PCR primers directed at mutant sites included in CYP2C9 and VKORC1 genes and closely related to metabolism of warfarin are designed, and the genotypes of to-be-detected genes are determined by reading the lengths of multiplex PCR products with naked eyes via common gel electrophoresis, so the dosage of warfarin in medication is guided and individualized medication of patients is realized; and the detection kit and detection method have the advantages of rapidness, accuracy, low cost, etc.

Description

technical field [0001] The invention relates to the technical fields of molecular biology and nucleic acid detection, in particular to a warfarin individualized drug detection kit and a detection method thereof. Background technique [0002] Warfarin is currently a coumarin-based oral anticoagulant drug widely used in clinical practice, which can prevent the formation and development of thrombus, such as the treatment of thromboembolic phlebitis, rheumatic heart disease, artificial heart valve replacement surgery, hip joint fixation It can also be used as an adjuvant drug for patients with myocardial infarction. Although the anticoagulant effect of warfarin is stronger than that of other currently approved anticoagulant drugs, due to the great variability of the individual dose of warfarin, if the drug cannot be used according to individual differences, it will lead to serious anticoagulant deficiency or drug overdose Fatal hemorrhage is caused, which brings great challenge...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
Inventor 汪大为
Owner BEIJING JINQI BIOLOGICAL TECH CO LTD
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