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Kit and method for detecting polymorphism of CYP2C19 gene

A technology of CYP2C19 and gene polymorphism, applied in the field of fluorescent quantitative PCR, can solve the problems of low sensitivity and specificity of polymorphism, low sensitivity and specificity, long detection time, etc., to eliminate false positives and false negatives , avoid false positive or false negative, high accuracy effect

Active Publication Date: 2013-12-25
刘辉
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  • Description
  • Claims
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Problems solved by technology

[0006] In view of the problems in the prior art that the sensitivity and specificity of detecting CYP2C19 gene CYP2C19*2, CYP2C19*3 and CYP2C19*17 polymorphisms are not high, and the detection time is long, the purpose of the present invention is to provide a method for detecting polymorphisms of CYP2C19 genes. morphological kit
[0008] In order to solve the problems of low sensitivity and specificity and long detection time in detecting CYP2C19 gene CYP2C19*2, CYP2C19*3 and CYP2C19*17 polymorphisms in the prior art, the present invention adopts the following technical solutions:

Method used

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  • Kit and method for detecting polymorphism of CYP2C19 gene
  • Kit and method for detecting polymorphism of CYP2C19 gene
  • Kit and method for detecting polymorphism of CYP2C19 gene

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Embodiment 1

[0052] Embodiment 1. Preparation of kit

[0053] 1. Design of internal reference gene GAPDH and detection primers and fluorescent probes

[0054] The primers and fluorescent probes specific to the above-mentioned gene sequences were respectively designed according to the gene sequences. Among them, the GAPDH gene sequence and the CYP2C19 gene sequence were obtained from the National Center for Biotechnology Information Nucleic Acid Database (NCBI), and the GAPDH gene ID was 2597. The reference sequence The number is NG_007073.2, and also refer to SEQ ID NO: 13 in the sequence listing of the present invention; the CYP2C19 gene ID is 1557, and the reference sequence number is NG_008384.1. Primer5.0 primer design software was used to design the following primers: CYP2C19 gene CYP2C19*2 polymorphism specific upstream and downstream primers and the CYP2C19 gene CYP2C19*2 polymorphism specific Taqman fluorescent probe, CYP2C19 gene CYP2C19*3 The specific upstream and downstream pri...

Embodiment 2

[0081] Example 2. Detection of CYP2C19 gene polymorphism with the kit of Example 1

[0082] Take the results of random detection of peripheral blood samples from 30 subjects as an example.

[0083] The detection process for detecting CYP2C19*2 gene polymorphism, CYP2C19*3 gene polymorphism and CYP2C19*17 gene polymorphism of a certain subject with the kit of the present invention is as follows: first obtain the peripheral blood sample of the clinical subject , to quickly extract genomic DNA; secondly, first prepare the fluorescent quantitative PCR reaction solution of the internal reference gene GAPDH and the internal positive control sequence, and dilute the internal positive control sequence standard and the internal reference gene GAPDH standard to a copy number / mL of 1.0x10 3 , 1.0x10 4 , 1.0x10 5 and 1.0x10 6 , make the standard curve of the internal positive control sequence standard and the standard curve of the internal reference gene GAPDH standard respectively; th...

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Abstract

The invention relates to a kit and a method for detecting polymorphism of CYP2C19 gene, and belongs to the field of fluorogenic quantitive PCR (Polymerase Chain Reaction). The kit comprises a detection primer and a fluorescent probe, which comprise at least one group of specific primer and specific Taqman fluorescent probe of CYP2C19 gene CYP2C19*2 polymorphism, specific primer and specific Taqman fluorescent probe of CYP2C19 gene CYP2C19*3 polymorphism, and specific primer and specific Taqman fluorescent probe of CYP2C19 gene CYP2C19*17 polymorphism. The kit is adopted for detecting the CYP2C19 gene, the sensitivity and specificity are both obviously improved, the detection time is short and the predication of medicament dosage is facilitated.

Description

technical field [0001] The invention relates to the field of fluorescent quantitative PCR, in particular to a kit and method for detecting CYP2C19 gene polymorphism. Background technique [0002] The CYP2C19 gene is an important drug-metabolizing enzyme that plays a key role in drug response. This is because there are significant individual differences in the activity of the CYP2C19 gene, manifested as genetic polymorphisms, resulting in individual differences in blood drug concentrations , the study of its genetic polymorphism will provide guidance for clinically determining the optimal therapeutic dose, formulating a rational drug regimen, maximizing the curative effect, avoiding and reducing adverse reactions. [0003] In the CYP2C19 gene, the distribution frequencies of CYP2C19*2 polymorphism and CYP2C19*3 polymorphism in the Chinese population are 39.9% and 6.1%, respectively, which lead to the loss of activity of the protein encoded by CYP2C19, which leads to drug meta...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N21/64
Inventor 刘辉
Owner 刘辉
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