Method for detecting BRAF (block repeat active flag) gene mutation accurately based on probe fusion technology

A technical detection and melting technology, applied in biochemical equipment and methods, microbial determination/inspection, fluorescence/phosphorescence, etc., can solve the problems of inaccurate detection and low sensitivity, and achieve low quality requirements, high sensitivity, and high sensitivity. Effect

Active Publication Date: 2015-07-15
陕西佰美医学检验有限公司
View PDF5 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to overcome the shortcomings of low sensitivity and inaccurate detection of the traditional probe melting curve method, the present invention proposes a method for accurately detecting BRAF gene mutations based on probe melting technology, which not only saves consumables, but also has the characteristics of high sensitivity and high specificity , the test results are more reliable

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
  • Method for detecting BRAF (block repeat active flag) gene mutation accurately based on probe fusion technology
  • Method for detecting BRAF (block repeat active flag) gene mutation accurately based on probe fusion technology
  • Method for detecting BRAF (block repeat active flag) gene mutation accurately based on probe fusion technology

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0033] The present invention combines molecular beacon probes with extension-blocking primers, and uses a polymerase without 5'→3' exonuclease activity to rapidly detect BRAF mutations in vitro, which not only saves consumables, but also has high sensitivity and high specificity The characteristics make the test results more reliable.

[0034] Extension-Refractory primers (Extension-Refractory primers) are primers whose self-extension products can form a "neck loop" structure; in addition to the normal primer sequence, the 5' end of the extension-refractory primer contains an additional base sequence, which The segment sequence can be complementary to a base sequence including the mutation site in the single strand of the product obtained by its own extension, forming a "neck ring" structure; due to the difference in sequence between the wild-type and mutant templates, the formed "neck ring" "There is a significant difference in structural strength (Tm value).

[0035]The pre...

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 provides a method for detecting BRAF (block repeat active flag) gene mutation accurately based on a probe fusion technology. Through the use of the method, not only consumable items are saved, but also characteristics of high sensitivity and specificity are realized and the detection result is more reliable. The method comprises the following steps of (1) design and synthesis of a molecular beacon probe and a template amplification primer, wherein a designed upstream primer (27nt) is 5'-TGTTTTCCTTTACTTACTACACCTCAG-3', an extension refractory primer (43nt) is 5'-acGCTACAGTGAAATCTCGGACAACTGTTCAAACTGATGGGAC-3', and the molecular beacon probe (35nt) is FAM-ccgcaTCGAGATTTCACTGTAGCTAGACCAtgcgg-BHQ2; (2) extraction of a genomic DNA (deoxyribonucleic acid) of a tested sample; (3) preparation of a reaction system; and (4) reaction of PCR (polymerase chain reaction).

Description

technical field [0001] The invention relates to a method for detecting BRAF gene mutation based on probe melting technology. Background technique [0002] Gene mutation refers to the change of base composition or arrangement sequence in the structure and function of genomic DNA molecules, mainly including base substitution and fragment insertion and deletion, and is one of the important causes of genotype diseases. Gene mutation analysis plays a very important role in biomedical research, especially in genotypic disease diagnosis and pathological research. The protein encoded by the BRAF gene is involved in the cell signal transduction pathway mediated by the epidermal growth factor receptor (EGFR), affecting cell proliferation, growth and metastasis; the protein encoded by the normal BRAF gene is activated after receiving upstream signal activation, and After the signal is transmitted to the downstream cytokines, the inactive state will be restored; while the protein encod...

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 Patents(China)
IPC IPC(8): C12Q1/68G01N21/64
Inventor 戴鹏高陈超刘金辉焦维丽邹晖
Owner 陕西佰美医学检验有限公司
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