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

Fluorescence immunity chromatography test strip for escherichia coli O157:H7 detection

A technique of fluorescence immunochromatography and O157, which is applied in the detection field to achieve the effects of fast detection speed, high sensitivity and strong specificity

Inactive Publication Date: 2013-10-30
HUAZHONG AGRI UNIV
View PDF9 Cites 15 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are no reports on the detection of E. coli O157:H7 using quantum dot-labeled immunochromatographic test strips

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
  • Fluorescence immunity chromatography test strip for escherichia coli O157:H7 detection
  • Fluorescence immunity chromatography test strip for escherichia coli O157:H7 detection
  • Fluorescence immunity chromatography test strip for escherichia coli O157:H7 detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Preparation and condition optimization of water-soluble core-shell quantum dots

[0030] CdTe / CdS core-shell quantum dots were synthesized in aqueous solution by using an inorganic aqueous phase synthesis method with mercaptopropionic acid as a stabilizer. The preparation conditions of CdTe / CdS quantum dots were optimized through single factor experiments, and the optimal level of each influencing factor was obtained: Cd 2+ with Te 2- The molar ratio is 5:1, the stabilizer MPA and Cd 2+ The molar ratio of CdTe is 2.5:1, the pH of the reaction system is 9, the reaction temperature is 95°C, the reflux time of CdTe core is 1h; the reflux time of CdS shell is 1h. Under these conditions, the fluorescence intensity of the synthesized CdTe / CdS is 726.41, which is 50.10% higher than the initial condition. The particle size of the quantum dots is about 3.0nm, and the dispersion is good. Compared with CdTe quantum dots, the core-shell structure The fluorescence inten...

Embodiment 2

[0039] Example 2 Labeling of E. coli O157:H7 rabbit polyclonal antibody and preparation of fluorescent immunochromatographic test strips

[0040] 1) Conditions for quantum dot-labeled Escherichia coli O157:H7 rabbit polyclonal antibody

[0041] Covalently coupled with EDC·HCL and NHS as coupling agent, take 100 μL of quantum dots, add 40 μL 0.1 mol / L EDC·HCl and 20 μL 0.1 mol / L NHS respectively, supplement pH 7.4, 0.01 mol / L PBS to The total volume was 800 μL, activated at room temperature for 15 minutes, then 80 μL of rabbit IgG with a concentration of 1 mg / mL was added, and the mixture was reacted in a shaker at 37 °C in the dark for 1.5 h, and finally the reaction was terminated with 100 μL of 1 mg / mL glycine, placed in overnight at 4°C.

[0042] The reaction mixture was repeatedly ultrafiltered with a 100k ultrafiltration membrane to remove the CdTe / CdS quantum dots that were not coupled to the Escherichia coli O157:H7 polyclonal antibody, and the CdTe / CdS-antibody conjug...

Embodiment 3

[0048] The test of embodiment 3 samples and result judgment

[0049] Add the sample liquid to the sample pad dropwise, or insert one end of the sample pad into the sample liquid. After the immunochromatography is finished, irradiate the result under the ultraviolet light. If the detection line (C line) and the quality control line (T line) appear at the same time A yellow fluorescent band is a positive result; if there is a fluorescent band on the C line and no fluorescent band on the T line, it is a negative result; if neither the C line nor the T line has a fluorescent band, or the T line has a fluorescent band but the C line does not. If the fluorescent band appears, it is judged that the test strip is invalid ( Figure 8 ).

[0050] 1) Detection limit: heat inactivate the pure culture of Escherichia coli O157:H7 and dilute to different concentrations respectively, add dropwise to the sample pad, carry out chromatographic reaction, irradiate the results under ultraviolet l...

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 sizeaaaaaaaaaa
Login to View More

Abstract

The invention discloses a fluorescence immunity chromatography test strip for escherichia coli O157:H7 detection. The test strip is prepared by a method comprising the following steps: 1) preparing water-soluble CdTe / CdS quantum dots with a core-shell structure; 2) marking escherichia coli O157:H7 rabbit polyclonal antibodies with the CdTe / CdS quantum dots; and 3) coating the escherichia coli O157:H7 marked rabbit polyclonal antibodies on a glass cellulose membrane to form a combination pad; respectively coating the escherichia coli O157:H7 polyclonal antibodies and goat anti-rabbit IgG on a cellulose nitrate membrane to form a detection line and a quality control line; and assembling a sample pad, the combination pad, the cellulose nitrate membrane and a piece of absorbent paper on a pvc base plate to form the fluorescence immunity chromatography test strip. The fluorescence immunity chromatography test strip has advantages of fast detection, high sensitivity, strong singularity and good stability in escherichia coli O157:H7 detection.

Description

technical field [0001] The invention belongs to the technical field of detection, and relates to a fluorescent immunochromatographic test strip for detecting Escherichia coli O157:H7 and an application thereof. Background technique [0002] Escherichia coli O157:H7 is an enteropathogenic bacterium that infects humans mainly through ingestion of contaminated food or water, or contact with carrier animals, and can cause death in severe cases. Escherichia coli O157:H7 infection has gradually become an important public health problem threatening human health, and the establishment of rapid and sensitive detection methods is the key to the prevention and control of Escherichia coli O157:H7 infection. [0003] Quantum dots are a new type of nanomaterial with excellent fluorescence properties. Compared with other immunological analysis methods, the quantum dot-based immunoassay method has great advantages in specificity, sensitivity and accuracy, and has broad application prospect...

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): G01N33/569
Inventor 郭爱玲苏靖张隽娴刘凤燕吕品田马美湖
Owner HUAZHONG AGRI 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