Unlock instant, AI-driven research and patent intelligence for your innovation.

Optimization method of PIK3CA gene E545K mutation digital PCR detection system and detection product

An optimized method and digital technology, applied in biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of low cfDNA content, insensitive and accurate detection of plasma free DNA mutation information, etc., and achieve accurate results

Pending Publication Date: 2019-02-01
上海赛安生物医药科技股份有限公司
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the content of ctDNA in cfDNA is extremely low, less than 1%, and traditional detection methods are not sensitive and accurate in detecting the mutation information contained in plasma cell-free DNA

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
  • Optimization method of PIK3CA gene E545K mutation digital PCR detection system and detection product
  • Optimization method of PIK3CA gene E545K mutation digital PCR detection system and detection product
  • Optimization method of PIK3CA gene E545K mutation digital PCR detection system and detection product

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] The PIK3CA gene E545K mutation digital PCR detection kit of this embodiment includes an upstream primer (PIK3CA-E545K-F), a downstream primer (PIK3CA-E545K-R), a wild-type probe (PIK3CA-E545K-wt) and a mutant probe (PIK3CA-E545K-mt), normal human gDNA after digestion and mutant plasmid after digestion.

[0034] The primers and probes are self-designed and optimized through multiple combinations. The primer probes include the homologous region of the mutant fragment inserted in the mutant plasmid. The primers and probes were synthesized by Shanghai Bailige Biotechnology Co., Ltd. The nucleotide sequences of the primers and probes are shown in Table 1.

[0035] Table 1 Primer probe sequence list

[0036] name

Sequence(5’—3’)

Seq No.

PIK3CA-E545K-F

TCAAAGCAATTTCTACACGAGATCCT

1

PIK3CA-E545K-R

GCACTTACCTGTGACTCCATAGAAA

2

PIK3CA-E545K-wt

CTCTCTGAAATCACTGAGCAG

3

PIK3CA-E545K-mt

CTCTGAAATCACTAAGCAG

4

[0037] The 5'end of the wild-type probe (PIK3CA-E545K-wt) is the VI...

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

The invention relates to an optimization method of a PIK3CA gene E545K mutation digital PCR detection system, wherein the detection system includes upstream primers, downstream primers, wild-type probes, mutant-type probes, a wild-type template and a mutant-type template; the wild-type template is normal human gDNA after enzyme digestion, and the mutant-type template is a mutant plasmid inserted with a PIK3CA gene E545K mutant fragment after enzyme digestion; a standard substance is prepared from the mutant-type template and the wild-type template according to a certain proportion of copy number; reaction is performed with a medium mutation frequency standard substance as a template by digital PCR, a data statistical graph is prepared according to the reaction data, and a wild-type fluorescent region and a mutant-type fluorescent region are selected. Reaction is performed with the wild-type template as the template by digital PCR, and the background threshold value is determined. A detection product optimized by the optimization method of the PIK3CA gene E545K mutation digital PCR detection system has higher accuracy degree.

Description

Technical field [0001] The invention relates to a digital PCR detection product for detecting E545K mutation of human PIK3CA gene and an optimization method thereof, and belongs to the field of biotechnology. Background technique [0002] Phosphatidylinositol-3-kinases (PI3Ks) are lipid kinases that are involved in the regulation of cell survival, movement, adhesion, and apoptosis. PIK3 is a heterodimer composed of the regulatory subunit p85 and the catalytic subunit p110. The p110α catalytic subunit of PI3Ks is encoded by the PIK3CA gene and is located at 3q26.3 and is 34kb long. PI3Ks are activated by growth factor receptor tyrosine kinases such as epidermal growth factor receptor (EGFR), insulin receptor, etc., and act as a signal transmitter inside the cell. The activated PI3Ks catalyze the phosphorylation of phosphatidylinositol 4 phosphate (PI-4-P) and phosphatidylinositol 4,5-bisphosphate (PI-4,5-P2) into phosphatidylinositol 3,4-bisphosphate ( PI-3,4-P2) and phosphatidy...

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): C12Q1/6886C12Q1/6858C12Q1/6851
CPCC12Q1/6886C12Q1/6851C12Q1/6858C12Q2600/156C12Q2531/113C12Q2563/107C12Q2545/114
Inventor 赵新泰王明潘文健
Owner 上海赛安生物医药科技股份有限公司
Features
  • R&D
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

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

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com