Supercharge Your Innovation With Domain-Expert AI Agents!

Method for visually detecting nucleic acid based on enzyme catalysis circulation and molybdenum disulfide adsorption mediation

A molybdenum disulfide and nucleic acid technology, applied in the field of colorimetric reagent research and nucleic acid detection, can solve the problems of complex operation and low sensitivity, and achieve the effects of good selectivity, improved detection sensitivity and simple operation

Active Publication Date: 2019-12-20
NANJING UNIV OF POSTS & TELECOMM
View PDF4 Cites 7 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Purpose of the invention: Aiming at the problems of low sensitivity and complicated operation in the visual nucleic acid detection in the prior art, the present invention provides a method for visual nucleic acid detection mediated by enzyme-catalyzed cycle and molybdenum disulfide adsorption

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 visually detecting nucleic acid based on enzyme catalysis circulation and molybdenum disulfide adsorption mediation
  • Method for visually detecting nucleic acid based on enzyme catalysis circulation and molybdenum disulfide adsorption mediation
  • Method for visually detecting nucleic acid based on enzyme catalysis circulation and molybdenum disulfide adsorption mediation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Nanogold-capture probe preparation:

[0040] (1) Mix the polyA-modified target nucleic acid capture sequence with 15nm gold nanoparticles at a molar ratio of 200:1 (such as a final concentration of 600nM DNA: 3nM gold nanoparticles), and shake gently overnight at room temperature at 25°C. The capture sequence is: SEQ ID NO.15'-AAAAAAAAAAAAAAAAAAAAACTCTCTAAAATCACT-3'(capture sequence modified by polyA); in the above steps, the target nucleic acid capture sequence modified by sulfhydryl group can also be used, SEQ ID NO.2: 5'-SH-CTCTCTAAAATCACT-3'(capture sequence modified by sulfhydryl group capture sequence), the follow-up effect is consistent with the polyA modified target nucleic acid capture sequence.

[0041] (2) Add 1M PBS (100mM PB, 1M NaCl, pH=7.4) to the solution of step (1) by volume ratio, add once every 30min, and add in five times, so that the final concentration of the mixed solution is 0.1M PBS ( 10 mM PB, 0.1 M NaCl, pH=7.4), and continue shaking overnig...

Embodiment 2

[0046] Take 100 μL of the nano-gold capture probe solution prepared in Example 1 (the final concentration of the probe is 6 nM), and add the target nucleic acid with a final concentration of 200 nM to it respectively. The sequence of the target nucleic acid is: SEQ ID NO.3: 5' -AGTGATTTTAGAGAGAG-3', after hybridization at room temperature for 10 minutes, exonuclease III at a final concentration of 0.1 U / μL was added, and reacted at 37°C for 30 minutes. Then add molybdenum disulfide nanosheets (size 200nm×200nm) with a final concentration of 15 μg / mL, and the transmission electron microscope image of molybdenum disulfide is shown in figure 2 ), let stand to react for 70min, take the supernatant to observe the color change of the supernatant with naked eyes, and measure the absorbance value change of the supernatant by an ultraviolet spectrophotometer.

Embodiment 3

[0048] The method of embodiment 3 is the same as that of embodiment 2, except that the particle diameter of gold nanoparticles is 10nm; the hybridization time between target nucleic acid and capture probe is 30min; the final concentration of exonuclease III is 0.05U / μL, The shearing time is 90min; the size of the molybdenum disulfide nanosheets is 100nm*100nm, and the final concentration is 18μg / mL; the static reaction time is 10min.

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
particle diameteraaaaaaaaaa
particle diameteraaaaaaaaaa
Login to View More

Abstract

The invention discloses a method for visually detecting nucleic acid based on enzyme catalysis circulation and molybdenum disulfide mediated adsorption. The method comprises the following steps: assembling a sulfydryl or polyA-modified capture sequence on the surfaces of gold nanoparticles to construct a nanogold capture probe; adding the capture probe into a to-be-detected system containing target nucleic acid, and carrying out cyclic shearing reaction after the target nucleic acid and the capture probe are hybridized; adding molybdenum disulfide nanosheets into a system after the shearing reaction, and performing standing for reaction; and taking a supernate of the system after the reaction in the previous step, observing the color change of the supernate by naked eyes, and measuring thelight absorption value change of the supernate by an ultraviolet spectrophotometer. Based on the enzyme circulation reaction triggered by the target nucleic acid, the method utilizes the adsorption of molybdenum disulfide and the nanogold probe to output signals, and can realize the on-site in-situ rapid detection of the target nucleic acid through naked eye observation or colorimetric spectrum.The method is simple to operate, mild in reaction condition, high in sensitivity and good in selectivity, and has a wide application prospect.

Description

technical field [0001] The invention belongs to the field of colorimetric reagent research and nucleic acid detection, and in particular relates to a method for visual detection of nucleic acid based on enzyme catalysis cycle and molybdenum disulfide adsorption-mediated visualization. Background technique [0002] In the past few years, nucleic acid detection has received high attention due to its important applications in medical research and diagnosis, as well as food and drug industry monitoring. Polymerase chain reaction (PCR) is the classic technology for nucleic acid detection. However, PCR needs to design complex primers for different targets, the steps are cumbersome, the cost is high, and it cannot meet the needs of clinical detection. The development of nano-biosensing technology provides a new opportunity for nucleic acid detection, usually by introducing a capture sequence complementary to the target nucleic acid in the nano-sensing system for nucleic acid recog...

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): C12Q1/6834
CPCC12Q1/6834C12Q2521/319C12Q2563/137C12Q2563/155C12Q2565/113C12Q2565/519
Inventor 汪联辉朱煜朱丹赵东霞晁洁
Owner NANJING UNIV OF POSTS & TELECOMM
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