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

Primer group, kit and method for detecting expression level of imatinib related gene C-kit

A technology of expression level and kit, which is applied in the field of primer set for the detection of expression level of imatinib-related gene C-kit, can solve the problems of various reagents, cumbersome experimental process, and great subjective influence on reading results, and achieve detection Accurate and reliable results, high detection sensitivity, and the effect of reducing medical costs

Inactive Publication Date: 2018-04-03
广东大臻医学检验所有限公司
View PDF2 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In order to solve the technical problems that the experimental process is too cumbersome in the detection of clinical biomarkers by immunohistochemistry, a wide variety of reagents are required, and the reading of the results is greatly influenced by the subjectivity, the present invention provides a method with simple operation, short detection time and high detection sensitivity. Primer set, kit and method for detection of expression level of imatinib-related gene C-kit with good specificity

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 group, kit and method for detecting expression level of imatinib related gene C-kit
  • Primer group, kit and method for detecting expression level of imatinib related gene C-kit
  • Primer group, kit and method for detecting expression level of imatinib related gene C-kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Embodiment 1: the preparation of kit

[0049] 1. Design and synthesis of primers and probes

[0050] For the C-kit gene in the human genome (for the sequence, refer to the human genome sequence published in the NCBI database), use Primer Premier 3.0 and Methyl Primer Express v1.0 software to design specific primers and probes, respectively, and use 5'-FAM as the probe and 3'-TRAMAMGB tag.

[0051] Specific primer and probe sequences:

[0052]

[0053]2. Selection of reference substance

[0054] The positive control substance is a solution containing C-kit gene; the negative control substance is a solution without C-kit gene; the blank control substance is ultrapure water.

[0055] 3. Composition of PCR reaction solution

[0056] Including the PCR reaction solution containing the above-mentioned specific primers and probes, the PCR reaction solution includes the specific primers and probes for detecting the C-kit gene and the reference gene Actin; it also includes...

Embodiment 2

[0060] Example 2: The method for detecting the expression level of genes related to individualized drug use of clinical imatinib using the above kit:

[0061] 1. Biological samples

[0062] The biological materials used in the present invention are all from Guangdong Dazhen Medical Laboratory. 100 remaining tissue samples tested at Guangdong Dazhen Medical Laboratory during January-December 2016.

[0063] 2. Take the sample to be tested to extract mRNA, reverse transcribe the sample mRNA to obtain cDNA, and use cDNA as a template:

[0064] Sample mRNA extraction kits were QIAGEN miRNeasy FFPE Kit (paraffin-embedded tissue sample RNA extraction) (QIAGEN Biotechnology Co., Ltd. (Shanghai)) and QIAGEN RNeasy Mini Kit (fresh tissue sample RNA extraction) (QIAGEN Biotechnology Technology Co., Ltd. (Shanghai));

[0065] The sample mRNA reverse transcription kit was Takara-PrimeScriptTM RT reagent Kit (Bao Bioengineering (Dalian) Co., Ltd.).

[0066] 1) Take qualified clinical te...

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

PropertyMeasurementUnit
correlation coefficientaaaaaaaaaa
Login to View More

Abstract

The invention relates to a primer group for detecting the expression level of an imatinib related gene C-kit. The primer group comprises a specific primer and a probe aiming a C-kit gene and a reference gene Actin. The invention also relates to a kit for detecting the expression level of the imatinib related gene C-kit. The kit comprises a PCR reaction liquid; the PCR reaction liquid comprises a specific primer and a probe for detecting the C-kit gene and the reference gene, and also comprises PCR buffer, dNTP, HS Taq enzyme and ultrapure water. The invention also relates to a method for detecting the expression level of the imatinib related gene C-kit. The kit and the detection method thereof have the advantages of simplicity in operation, short detection time, high detection sensitivity,high specificity and the like, and are particularly suitable for popularization and application in clinical inspection work.

Description

technical field [0001] The invention relates to the technical field of in vitro diagnosis, in particular to a primer set, a kit and a method for detecting the expression level of an imatinib-related gene C-kit. Background technique [0002] C-kit (V-kit hardy-zuckerman 4 feline sarcoma viral oncogene homolog) protein is a member of the type III transmembrane receptor tyrosine kinase family encoded by the proto-oncogene C-kit. It consists of an intracellular tyrosine kinase domain, The transmembrane region and the extracellular region with the ligand binding site. After binding with its ligand, it can activate its own tyrosine protease activity and activate downstream signal transduction pathways through a series of reactions, such as: PI3K pathway, JAK-STAT pathway, Ras-Erk pathway, SFK pathway, phospholipase C pathway, etc. Ultimately activates transcription factors in cells, thereby regulating gene expression, controlling cell growth and proliferation. Studies have found...

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
Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/6886C12Q2600/106C12Q2600/158C12Q2561/101C12Q2563/107C12Q2545/101
Inventor 不公告发明人
Owner 广东大臻医学检验所有限公司
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