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

A kind of multiple nucleic acid exponential amplification probe and its application in tumor multi-target detection

An exponential amplification, multiple nucleic acid technology, applied in the field of tumor detection kits and multiple nucleic acid exponential amplification probes, can solve the problems of difficult to achieve multiple nucleic acid exponential amplification, poor specificity, low sensitivity, etc., and achieve good clinical practical value. With the application prospect, high sensitivity, the effect of eliminating interference

Active Publication Date: 2022-02-18
XI AN JIAOTONG UNIV
View PDF8 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The object of the present invention is to provide a multiple nucleic acid exponential amplification probe and its tumor detection kit and application to overcome the low sensitivity, poor specificity and difficulty in realizing multiple nucleic acid exponential amplification in existing nucleic acid detection methods Shortcomings

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
  • A kind of multiple nucleic acid exponential amplification probe and its application in tumor multi-target detection
  • A kind of multiple nucleic acid exponential amplification probe and its application in tumor multi-target detection
  • A kind of multiple nucleic acid exponential amplification probe and its application in tumor multi-target detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Select the mRNA Tk1 in MCF-7 cells as the tumor target nucleic acid, synthesize oligonucleotides with the same sequence as Tk1 mRNA to simulate the target to be tested, the reaction concentration of TK1 padlock is 500nM, the reaction concentration of target nucleic acid Tk1 to be tested is 100nM, and react at 65°C , 5min, cool at room temperature for 1h; add 1μL 100U / μL T4 ligase, react at 16°C for 2h, 65°C for 10min, to ligate the TK1 padlock into a circular template. Take 4 μL of TK1 padlock connected into a circle (reaction concentration 100 nM) for amplification, the amplification reaction system is 20 μL in total, including 4 μL of the TK1 padlock connected into a circle above, 1 μL of 20 μM forward amplification primer, 1 μL of 20 μM reverse amplification primer, 2 μL 10× isothermal amplification buffer; 1 μL 10mM dNTPs; 5 μL 5M betaine; 1 μL 100mM Mg 2+ ; 1 μL 10 μM fluorescent probe; 2 μL H 2 O; 2μL 2U / μL Bst DNA polymerase to react at 65°C for 3h, then at 80°C...

Embodiment 2

[0052] Taking MCF-7 as an example, a variety of tumor target nucleic acids with important clinical significance in MCF-7 were selected for detection (Tk1, PFN1, GAPDH). Ligation reaction, amplification reaction experimental process as mentioned above, image 3 (a) shows the real-time fluorescence values ​​detected by Cy5 as a signal probe when different concentrations of Tk1 are exponentially amplified for 1 hour. The first red column from the left is the blank control, and the second red column is after 20nM Tk1 amplification The obtained fluorescence value, the third red column is the fluorescence value obtained after 200nM Tk1 amplification, it can be seen from the figure that the higher the concentration of Tk1, the higher the fluorescence value obtained by amplification, and the fluorescence value of the blank control is significantly lower Fluorescence value in the presence of target. (b) The picture shows the real-time fluorescence value detected by Texas Red as a sig...

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 multiple nucleic acid exponential amplification probe and its application in tumor multi-target detection, belonging to the technical field of gene detection. Only the sequence of the recognition region hybridized with the target nucleic acid is different among the different recognition probes for the target nucleic acid, and the rest of the positions are common sequences. All kinds of circular templates formed by target-specific ligation can achieve exponential amplification through a pair of universal amplification primers, avoiding the interference of different types of multiple primers or high concentrations on the amplification system, and eliminating the non-specific amplification of the system interference. At the same time, combining different fluorescent color combinations to mark the coding sequence of the amplified product can improve the information collection throughput of a single sample, and can accurately obtain multiple tumor marker nucleic acid information, providing new ideas for early diagnosis, prognosis evaluation and treatment monitoring of tumors.

Description

technical field [0001] The invention belongs to the technical field of gene detection, and relates to a multiple nucleic acid exponential amplification probe, a tumor detection kit and an application based thereon. Background technique [0002] The occurrence and development of tumors is a highly complex event involving multiple molecules and multiple pathways. Comprehensive and accurate detection and determination of tumors requires simultaneous detection of multiple tumor marker nucleic acids. [0003] Most of the existing nucleic acid detection methods have low sensitivity and poor detection accuracy of low-abundance targets; at the same time, due to the single form of fluorescent signal and the presence of base bias and cross-interference between target-specific primers, it is difficult to achieve simultaneous detection of multiple targets in the same system . Contents of the invention [0004] The object of the present invention is to provide a multiple nucleic acid ...

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): C12Q1/6886C12Q1/6844C12N15/11
CPCC12Q1/6886C12Q1/6844C12Q2600/16
Inventor 赵永席赵越付有兰
Owner XI AN JIAOTONG UNIV
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