Kit for detecting B-raf gene V600E mutation, and detection method thereof

A b-raf, human detection technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve problems such as reducing PCR efficiency, and achieve the effect of good detection sensitivity and specificity, and economic detection.

Inactive Publication Date: 2018-07-13
天津安必森生物技术有限公司
View PDF0 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the above two methods reduce the efficiency of PCR amplific

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
  • Kit for detecting B-raf gene V600E mutation, and detection method thereof
  • Kit for detecting B-raf gene V600E mutation, and detection method thereof
  • Kit for detecting B-raf gene V600E mutation, and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0035] The present invention is specifically described below in conjunction with accompanying drawing,

[0036] See attached figure 1 , the test kit for detecting B-raf gene mutation of the present invention, for the first time combined with ARMS-PCR fluorescent PCR probe technology, competitive inhibition Blocker technology, oligonucleotide modification technology and TspR I restriction enzyme digestion technology, The advantage of this design step is that the sensitivity and efficiency of detection can be satisfied to the greatest extent.

[0037] See attached figure 2 , the kit for detecting B-raf gene mutation of the present invention introduces a two-base mutation mechanism at the 3rd and 5th positions of the upstream primer of ARMS-PCR for the first time. Introducing a single base mutation at the 3' end of the upstream primer of ARMS-PCR is a routine method for ARMS-PCR; introducing an additional mutation at the third position is an improvement of the ARMS-PCR techniq...

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 discloses a kit for detecting B-raf gene V600E mutation, and a detection method thereof. The kit comprises B-raf gene V600E mutation detection mixed liquid, wherein the B-raf gene V600Emutation detection mixed liquid comprises a B-raf gene V600E mutation detection specific primer pair, a B-raf gene specific probe, a competitive inhibition nucleotide sequence, thermostability DNA (deoxyribonucleic acid) polymerase, thermostability restriction enzyme, reaction buffer solution and reaction enhancer. The kit has the beneficial effects that an ARMS-PCR (Amplification Refractory Mutation System-Polymerase Chain Reaction) fluorescence probe technology, a competitive inhibition Blocker technology, an oligonucleotide modification technology and a TspRI restriction enzyme technology are subjected to combined adaptation, mutation samples and wild samples can be more favorably distinguished so as to realize detection sensitivity and specificity better than similar technologies or products, B-raf gene V600E site mutation can be quickly, conveniently, sensitively and economically detected, and the kit is suitable for popularization application and industrialization.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a kit for detecting human B-raf gene V600E mutation and a detection method thereof. Background technique [0002] The B-raf gene is an oncogene, which encodes a serine / threonine-specific kinase, and is the most critical activator of the RAS / RAF / MEK / ERK / MAPK signaling pathway, involved in the regulation of cell growth, differentiation, and apoptosis. Studies have shown that in a variety of human malignant tumors, such as colorectal cancer, thyroid cancer, melanoma and lung cancer, there are different proportions of B-raf mutations. Among them, 80-90% of the mutations are V600E (1799T>A) mutations, which in turn lead to the phosphorylation of T598 and S601, resulting in abnormal protein expression, thereby affecting tumor progression. Studies have shown that patients with B-raf gene mutations are less effective in receiving EGFR-TKI drug therapy, and B-raf V600E mutation can...

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): C12Q1/6886C12Q1/686
CPCC12Q1/686C12Q1/6886C12Q2600/156C12Q2535/137C12Q2563/107C12Q2525/10C12Q2521/301
Inventor 张亮钟建方平科黄明
Owner 天津安必森生物技术有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap