Nanometer biological membrane for quickly detecting escherichia coli and preparation method thereof

A technology of biofilm and nanofiber membrane, which is applied in the field of nanobiofilm, can solve the problems of time-consuming, high detection limit of disease source samples, and long time-consuming, etc., and achieve the effect of simple operation, high sensitivity, and short detection time

Inactive Publication Date: 2012-06-27
CHANGSHA UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Abstract
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AI Technical Summary

Problems solved by technology

[0002] Routine microbial analysis requires targeted selection of medium to enrich the number of pathogens in the sample, so it takes 4-8 days from sample delivery to laboratory results, which takes a long time, and the reproduction of microorganisms often occurs within 1-2 days There may be an outbreak situation, so there is an urgent need for rapid diagnostic detection technologies that can significantly reduce analysis time, reduce usage costs, and effectively prevent or reduce foodborne disease outbreaks
At present, the rapid diagnosis and detection technology for foodborne diseases is mainly based on the lateral flow format (the lateral flow format) diagnosis of immunological assays. Although it is fast, the detection limit of the required pathogenic sample volume (0.1ml) is relatively high, about 4log cfu / ml, also need to enrich the number of pathogens, so time-consuming

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Preparation of nanobiofilms:

[0022] (1) Preparation of nanofiber membrane: electrospinning a nylon 6 formic acid solution with a mass concentration of 20% to form a nanofiber membrane; take the membrane and measure 100 fibers with an electron microscope, and the average diameter is 94.11nm-209.49nm.

[0023] (2) Synthesis of polyaniline-nylon nanofiber membrane: put the above nanofiber membrane into 0.40mol / L aniline hydrochloric acid solution at 40°C for 90 minutes, discard the reaction solution, and then add ammonium persulfate hydrochloric acid solution in equimolar ratio to aniline , polymerized at 0°C for 60 minutes, discarded the polymerization solution, rinsed until the washing solution was colorless, and dried at room temperature to obtain a polyaniline-nylon nanofiber membrane;

[0024] (3) labeling of antibodies; on the above-mentioned polyaniline-nylon nanofiber membrane, use 3% (w / v) glutaraldehyde as a cross-linking agent to couple 30mg / 100ml bovine serum...

Embodiment 2

[0028] Preparation of nanobiofilms:

[0029] (1) Preparation of nanofiber membrane: electrospinning a nylon 6 formic acid solution with a mass concentration of 16% to form a nanofiber membrane; take the membrane and measure 100 fibers with an electron microscope, and the average diameter is 94.11nm-209.49nm.

[0030] (2) Synthesis of polyaniline-nylon nanofiber membrane: put the above nanofiber membrane into 0.35mol / L aniline hydrochloric acid solution at 38°C for 85 minutes, discard the reaction solution, and then add ammonium persulfate hydrochloric acid solution in equimolar ratio to aniline , polymerized at 0°C for 45 minutes, discarded the polymerization solution, rinsed until the washing solution was colorless, and dried at room temperature to obtain a polyaniline-nylon nanofiber membrane;

[0031] (3) labeling of antibodies; on the above-mentioned polyaniline-nylon nanofiber membrane, use 2% (w / v) glutaraldehyde as a cross-linking agent to couple 15mg / 100ml bovine serum...

Embodiment 3

[0035] Preparation of nanobiofilms:

[0036] (1) Preparation of nanofiber membrane: electrospinning a nylon 6 formic acid solution with a mass concentration of 24% to form a nanofiber membrane; take the membrane and measure 100 fibers with an electron microscope, and the average diameter is 94.11nm-209.49nm.

[0037] (2) Synthesis of polyaniline-nylon nanofiber membrane: put the above nanofiber membrane into 0.45mol / L aniline hydrochloric acid solution at 43°C for 95 minutes, discard the reaction solution, and then add ammonium persulfate hydrochloric acid solution in equimolar ratio to aniline , polymerized at 0°C for 90 minutes, discarded the polymerization solution, rinsed until the washing solution was colorless, and dried at room temperature to obtain a polyaniline-nylon nanofiber membrane;

[0038] (3) labeling of antibodies; on the above-mentioned polyaniline-nylon nanofiber membrane, use 4% (w / v) glutaraldehyde as a cross-linking agent to couple 50mg / 100ml bovine serum...

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Abstract

The invention provides a nanometer biological membrane for quickly detecting escherichia coli and a preparation method thereof. The preparation method of the nanometer biological membrane comprises the following steps of: nanometer fiber membrane preparation, polyaniline-nylon nanometer fiber membrane synthesis and antibody labeling. The invention has the advantages that with the adoption of the nanometer biological membrane, the existence of the escherichia coli can be detected within 30 minutes, the detection is limited within 10-106 CFU / mL, and the nanometer biological membrane has higher sensitivity.

Description

technical field [0001] The invention relates to the field of nano-biofilms, in particular to a nano-biofilm for rapid detection of Escherichia coli and a preparation method thereof. Background technique [0002] Routine microbial analysis requires targeted selection of medium to enrich the number of pathogens in the sample, so it takes 4-8 days from sample delivery to laboratory results, which takes a long time, and the reproduction of microorganisms often occurs within 1-2 days There may be an outbreak situation, so there is an urgent need for rapid diagnostic detection technologies that can significantly reduce analysis time, reduce use costs, and effectively prevent or reduce foodborne disease outbreaks. At present, the rapid diagnosis and detection technology for foodborne diseases is mainly based on the lateral flow format (the lateral flow format) diagnosis of immunological assays. Although it is fast, the detection limit of the required pathogenic sample volume (0.1ml...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/531G01N27/12
Inventor 易翠平
Owner CHANGSHA UNIVERSITY OF SCIENCE AND TECHNOLOGY
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