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

Ratio-type electrochemical biosensor for eliminating LAMP background interference, construction method and application

A biosensor and construction method technology, applied in the field of ratiometric electrochemical biosensor construction, can solve problems such as difficulty in realization, achieve the effect of improving specificity and reducing false positive signals

Active Publication Date: 2020-04-24
SOUTHWEST UNIVERSITY
View PDF5 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

And this process is also extremely challenging, and it is difficult to achieve

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
  • Ratio-type electrochemical biosensor for eliminating LAMP background interference, construction method and application
  • Ratio-type electrochemical biosensor for eliminating LAMP background interference, construction method and application
  • Ratio-type electrochemical biosensor for eliminating LAMP background interference, construction method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] 1. Sequence design:

[0049] Taking HIV-specific marker DNA as the target (hDNA) analysis model and as the external primer F3 of the LAMP reaction, on the NUPACK website ( www.nupack.org ) to design all relevant base sequences based on hDNA. As shown in Table 1.

[0050] Table 1. DNA base sequence used in the present invention

[0051]

[0052]

[0053] Among them, the Mg formed by the hybridization of S* and C* 2+ -The hairpin configuration and base sequence of DNAzyme, MB-H1 and Fc-H2 are as follows figure 1 shown.

[0054] 2. A ratiometric electrochemical biosensor for eliminating LAMP background interference, its construction method and application, the specific steps are as follows:

[0055] (1) LAMP reaction ( figure 2 ): The target hDNA (F3) reacts with the template DNA, B3, BIP, FIP, polymerase Bst and dNTPs in a closed-tube PCR instrument, and the amplification product solution contains a large amount of H + (LAMP(H + ))

[0056] (2) LAMP (H +...

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
diameteraaaaaaaaaa
Login to View More

Abstract

The invention relates to a ratio-type electrochemical biosensor for eliminating LAMP background interference, a construction method and application. According to the method, a target object short-chain DNA is used as an LAMP amplification outer primer, the byproduct hydrogen ions induce i-motif conformation transformation, metal ions DNAzyme are assembled, cyclic shearing is carried out, and separation and purification of an LAMP complex system and products are achieved; two DNA hairpins marked with electroactive probes methylene blue and ferrocene respectively are triggered on the surface ofthe electrode for directional hybridization without any enzyme intervention, and when the two DNA hairpins are far away from and close to the surface of the electrode respectively, the current signalratio has high-sensitivity and high-specificity target concentration linear correlation. The electrode reaction of the established ratio-type sensor is more simplified, background interference and false positive signals are effectively reduced, the sensitivity is remarkably improved, and a new thought and a technical means are provided for electrochemical detection of DNA of various disease markers.

Description

technical field [0001] The invention belongs to the technical field of molecular biology and electrochemical biosensor, and relates to a construction method and application of a ratiometric electrochemical biosensor for eliminating LAMP background interference. Background technique [0002] As an important and efficient nucleic acid amplification technique, loop-mediated isothermal amplification (LAMP) can achieve 10 9 Replicate amplification. During the LAMP reaction, in addition to the main products of double-stranded DNA of different lengths, there are also a large amount of H + And pyrophosphoric acid, hydrolysis of pyrophosphoric acid further produces H + . Therefore, the pH of the LAMP reaction solution will vary depending on the target primer. [0003] Due to the advantages of mild reaction conditions, simple and fast operation, no need for special equipment, and high specificity, LAMP technology has been widely used in pathogen detection, clinical medicine, food ...

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): G01N27/327G01N27/30
CPCG01N27/3276G01N27/3278G01N27/308
Inventor 许文菊
Owner SOUTHWEST UNIVERSITY
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