Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Primer set for amplification of CYP2C19 gene, reagent for amplification of CYP2C19 gene comprising the same, and use of the same

A CYP2C19, gene amplification technology, applied in recombinant DNA technology, fermentation and other directions, can solve the problems of low reliability of analysis results, unpractical analysis, and a lot of labor, and achieves omitting pretreatment, shortening amplification reactions, and reducing labor and labor. cost effect

Inactive Publication Date: 2009-08-05
ARKRAY INC
View PDF4 Cites 8 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, when the genes encoding other isozymes are also amplified, for example, it becomes the analysis result of the specific polymorphism (CYP2C19*2 or CYP2C19*3) analysis of the CYP2C19 gene (Non-Patent Document 1 and Non-Patent Document 2) Causes of reduced reliability
Furthermore, in this way, a large amount of labor is required for analyzing a single sample, so there is also a problem that it is not practical to analyze a large number of samples.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Primer set for amplification of CYP2C19 gene, reagent for amplification of CYP2C19 gene comprising the same, and use of the same
  • Primer set for amplification of CYP2C19 gene, reagent for amplification of CYP2C19 gene comprising the same, and use of the same
  • Primer set for amplification of CYP2C19 gene, reagent for amplification of CYP2C19 gene comprising the same, and use of the same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0146] Blood was collected from 4 subjects using lithium heparin blood collection tubes (samples 1 to 4). 10 μL of the obtained blood was mixed with 90 μL of distilled water, and 10 μL of the mixed solution was mixed with 90 μL of distilled water. 10 μL of this mixed solution was added to 40 μL of a PCR reaction solution having the following composition, and PCR was performed using a thermal cycler. The conditions of PCR were as follows: after treatment at 95°C for 60 seconds, 1 cycle at 95°C for 1 second and 10 seconds at 54°C was repeated for 50 cycles, and then at 95°C for 1 second, and then Treatment at 40°C for 60 seconds. Next, the above-mentioned PCR reaction solution was heated from 40° C. to 95° C. at a rate of temperature increase of 1° C. / 3 seconds, and changes in fluorescence intensity over time were measured. The measurement wavelengths are 450-480 nm (detection of the fluorescent dye Pacific Blue), and 515-555 nm (detection of the fluorescent dye BODIPY FL). N...

Embodiment 2

[0169] Blood was collected from two subjects using EDTA blood collection tubes (Samples 1-2). 10 μL of the obtained blood was mixed with 70 μL of the following diluent A, and 10 μL of the mixed solution was mixed with 70 μL of the following diluent B. 10 μL of this mixture was heat-treated at 95° C. for 5 minutes, then added to 46 μL of a PCR reaction solution having the following composition, and PCR was performed using a thermal cycler. The conditions of PCR are as follows: after treating at 95°C for 60 seconds, repeating 50 cycles of 1 second at 95°C and 15 seconds at 64°C, and then treating at 95°C for 1 second, Treat at 40°C for 60 seconds. Next, the above-mentioned PCR reaction solution was heated from 40° C. to 75° C. at a rate of temperature increase of 1° C. / 3 seconds, and changes in fluorescence intensity over time were measured. The measurement wavelength is 515-555nm (detection of fluorescent dye BODIPY FL) and 585-700nm (detection of fluorescent dye TAMRA).

[...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

Disclosed is a primer set for amplifying a target region in CYP2C19 gene which contains a site to be detected by a gene amplification method, and which enables to amplify the region specifically. Two pairs of primer sets are used, each of which comprises a forward primer comprising a nucleotide sequence depicted in each of SEQ ID NOs:12 and 32 and a reverse primer comprising a nucleotide sequence depicted in each of SEQ ID NOs:22 and 48, respectively. These primer sets enable to amplify two regions each containing a site having each of two types of polymorphisms occurring in CYP2C19 gene (CYP2C19 gene*2, CYP2C19*3) simultaneously in a single reaction solution.

Description

technical field [0001] The present invention relates to a pair of primers for amplifying CYP2C19 gene, a reagent for amplifying CYP2C19 gene containing the primer pair and use thereof. Background technique [0002] Cytochrome P450 is an enzyme classified into a superfamily, and there are multiple subfamilies (for example, CYP1A, CYP1B, CYP2C, CYP2D, CYP2E, CYP3A, etc.). Among them, CYP2C19, which is an isozyme of the human CYP2C subfamily, is known as an enzyme involved in drug metabolism. Furthermore, mutations in the gene encoding CYP2C19 (CYP2C19 gene) have been demonstrated by enzyme-deficient persons (PMs) who have very low metabolism of a CYP2C19 substrate drug (antiepileptic agent), namely (S)-Mephenytoin (S-Mep) . CYP2C19*2 and CYP2C19*3 are known as important gene polymorphisms related to the PMs. The former is a mutation in which guanine (position 681) of exon 5 is changed to adenine, which causes a splicing defect and changes the translation initiation site of ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12Q1/68C12N15/09C12P19/34
Inventor 间岛智史吉永由纪
Owner ARKRAY INC
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com