Method for detecting relative gene polymorphism of warfarin personalized medication by using high-resolution melting curve analysis technology

A technology of gene polymorphism and high-resolution melting, which is applied in the field of detection of gene polymorphisms related to warfarin individualized medication, can solve the problems of primer design limitations, unsuitable detection, high cost, etc., and achieve fast detection speed, The effect of low cost and simple operation

Inactive Publication Date: 2014-01-22
上海中优医药高科技股份有限公司
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Problems solved by technology

This method has few steps, is not easy to pollute, and is convenient for typing large samples; however, it has certain restrictions on primer design and certain requirements on the sequence near the SNP, which is limited by the position and type of the mutated base
Direct DNA sequencing can directly provide accurate base information and is regarded as the gold standard for mutation detection, but it is time-consuming, laborious, expensive, and not suitable for detection of a large number of samples.

Method used

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  • Method for detecting relative gene polymorphism of warfarin personalized medication by using high-resolution melting curve analysis technology
  • Method for detecting relative gene polymorphism of warfarin personalized medication by using high-resolution melting curve analysis technology

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Embodiment

[0028] 1. The genomic DNA of the oral epithelial cells of the test subject was extracted by the silica gel adsorption method, and the concentration and purity of the DNA were detected by the electrophoresis gel imaging method, and the concentration of the sample to be tested was standardized to 10ng / ul;

[0029] The negative control DNA is considered qualified when it meets the following conditions: the OD value of A260 / A280 is between 1.8 and 2.0; the electrophoresis band is clear; the genotype of the CYP2C9 Arg144Cys (rs1799853) locus is identified by sequencing as CC;

[0030] Positive control 1 DNA is considered qualified when it meets the following conditions: OD value A260 / A280 is between 1.8-2.0; electrophoresis bands are clear; sequencing identifies CYP2C9 Arg144Cys (rs1799853) locus genotype as CT;

[0031] Positive control 2 DNA was considered qualified when it met the following conditions: OD value A260 / A280 was between 1.8-2.0; electrophoresis bands were clear; sequ...

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Abstract

The invention discloses a method for detecting relative gene polymorphism of warfarin personalized medication by using a high-resolution melting curve analysis technology. The method is characterized by comprising the following steps: adopting a silica gel adsorption method to extract genome DNA (Deoxyribose Nucleic Acid) of an oral epithelial cell / peripheral blood cell sample of a detected person; designing a detection primer containing a target site; carrying out site typing on a PCR (Polymerase Chain Reaction) amplified gene sequence by using the high-resolution melting curve (HRM) analysis technology. The method disclosed by the invention is high in sensitivity, good in specificity and fast in detection speed, and can be used for providing reference to a clinical medication dosage of warfarin, so as to realize reasonable and effective personalized medication.

Description

technical field [0001] The invention relates to a method for detecting gene polymorphisms related to individualized medicine of warfarin, and belongs to the technical field of molecular biology. Background technique [0002] Warfarin is a commonly used anticoagulant drug in clinical practice. It inhibits the formation of new thrombus by inhibiting the activation of coagulation factors, restricts the expansion and extension of thrombus, inhibits the occurrence of thrombus shedding and embolism, and is beneficial to the removal of thrombus. Clinically, it is mainly used for the treatment of atrial fibrillation, heart valve repair, recurrent stroke, deep vein thrombosis and pulmonary embolism. The pharmacology of warfarin is complex, and even small dose-response changes can lead to thrombosis or bleeding. The clinical dosage can vary by about 20 times. How to use warfarin correctly and monitor and adjust the dosage reasonably has become a difficult problem for clinicians. War...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/686C12Q2527/107C12Q2563/107
Inventor 吴迪傅咏南张奕王校毛丹丹卞雪莲
Owner 上海中优医药高科技股份有限公司
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