Imprinted thin film material for selectively separating drug-resistant bacillus as well as preparation and application thereof

An imprinted membrane and selective technology, which is applied in the direction of isolation of microorganisms, microorganism-based methods, biochemical equipment and methods, etc., can solve the problem of insufficient selectivity in the isolation of drug-resistant bacteria, the inability to effectively finely separate drug-resistant bacteria, and the elution of template bacteria To achieve the effect of promoting selective movement and active identification, effective and fine isolation of drug-resistant bacteria, and high selective isolation

Active Publication Date: 2020-11-10
HUAZHONG UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The invention solves the lack of selectivity for separating drug-resistant bacteria in the prior art, the difficulty in elution and identification of template bacteria, and the existing bacterial isolation method cannot effectively finely separate drug-resistant bacteria, and cannot meet the increasing requirements of existing microbial inspection and analysis. In order to solve the problems such as fine requirements, the present invention provides an imprinted film material for selectively isolating drug-resistant bacillus. The combination of induced active recognition, through the release of drug-resistant bacteria attractant and non-resistant bacteria repellent, promotes the selective movement and recognition of drug-resistant bacteria and avoids the interference of non-resistant bacteria, thereby improving the selection of drug-resistant bacteria sexual separation

Method used

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  • Imprinted thin film material for selectively separating drug-resistant bacillus as well as preparation and application thereof
  • Imprinted thin film material for selectively separating drug-resistant bacillus as well as preparation and application thereof
  • Imprinted thin film material for selectively separating drug-resistant bacillus as well as preparation and application thereof

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preparation example Construction

[0040] A method for preparing an imprinted film material for selectively isolating drug-resistant Escherichia coli of the present invention comprises the following steps:

[0041] Step 1: Dissolve polydimethylsiloxane (PDMS) and cross-linking agent in an appropriate amount of cyclohexane, apply the solution on a glass plate, and place it at 60-90°C for micro-curing for 3-5 minutes, wherein, The volume ratio of polydimethylsiloxane (PDMS) to the crosslinking agent is 1:0.08-0.15, and the volume of cyclohexane is 1-2 mL. At the same time, take an appropriate amount of drug-resistant Escherichia coli in the logarithmic growth phase and dilute to an appropriate concentration (OD 600 =0.06~0.08), and take 1~3mL of bacterial suspension dropwise onto a sterilized glass plate, let it stand at 3~6°C for 2~4 hours to allow the bacteria to settle, and then remove excess solvent in a sterile ventilated environment . Then the glass plate loaded with the template bacteria was pressed into...

Embodiment 1

[0047] Dissolve 2mL of polydimethylsiloxane (PDMS) and 0.2mL of cross-linking agent in 1mL of cyclohexane, and apply all of them evenly on a glass plate, and slightly cure at 80°C for 4min before use. At the same time, take an appropriate amount of drug-resistant Escherichia coli in the logarithmic growth phase, wash with PBS buffer solution with pH 7.4, and dilute to OD 600 = 0.07. Take 2 mL of the above bacterial suspension and drop it on a sterilized glass plate, let it stand at 4 °C for 2 h to allow the bacteria to settle, and then remove excess solvent in a sterile ventilated environment. A sterile glass plate coated with bacteria was pressed into the micro-cured PDMS surface, kept at 37.5 °C for 7 h, and further cured at 80 °C for 1 h. The imprinted film was peeled off from the glass plate, and then the template bacteria were washed and removed with deionized water under ultrasonic conditions. Place the imprinted film together with 10 mL of 0.1% methyltrichlorosilane s...

Embodiment 2

[0050] Dissolve 2mL of polydimethylsiloxane (PDMS) and 0.16mL of cross-linking agent in 2mL of cyclohexane, and apply all of them evenly on a glass plate, and slightly cure at 90°C for 3min before use. At the same time, take an appropriate amount of drug-resistant Escherichia coli in the logarithmic growth phase, wash with PBS buffer solution with pH 7.4, and dilute to OD 600 = 0.08. Take 1 mL of the above bacterial suspension and drop it on a sterilized glass plate, let it stand at 3°C ​​for 3 h to allow the bacteria to settle, and then remove excess solvent in a sterile ventilated environment. A sterile glass plate coated with bacteria was pressed into the above micro-cured PDMS surface, kept at 37.5 °C for 6 h, and further cured at 70 °C for 2 h. The imprinted film was peeled off from the glass plate, and then the template bacteria were washed and removed with deionized water under ultrasonic conditions. Place the imprinted film together with 5 mL of 0.15% methyltrichloro...

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Abstract

The invention discloses an imprinted film material for selectively separating drug-resistant bacilli as well as preparation and application thereof, and relates to the field of new materials. The method comprises the following steps: preparing a semi-cured film; preparing and dilution a drug-resistant bacillus suspension, dropwise adding the diluted drug-resistant bacillus suspension onto a sterilized substrate, performing standing treatment, and then removing the solvent in a sterile environment; pressing the substrate loaded with the drug-resistant bacilli into the semi-cured film so as to form prints on the semi-cured film with the drug-resistant bacilli as template molecules, and heating and completely curing the film; stripping the imprinted film from the substrate to obtain an imprinted film precursor, washing off drug-resistant bacilli as template molecules on the precursor, and carrying out silanization treatment on the surface of the precursor to enhance the hydrophilicity ofthe film; and forming a microporous structure into the imprinted film to obtain the imprinted film material for selectively separating the drug-resistant bacillus. On the basis of ensuring the recognition capability of the traditional bacterial imprinting material, the interference of non-drug-resistant bacteria can be avoided, and the drug-resistant bacteria are promoted to generate selective movement and recognition so as to achieve the purpose of selectively separating bacteria.

Description

technical field [0001] The present invention relates to the field of new materials, more specifically, to an imprinted film material for selectively isolating drug-resistant bacilli and its preparation and application, especially to an imprinted film material for selectively isolating drug-resistant bacilli based on bacterial chemotaxis preparation and application. Background technique [0002] In recent years, pathogens have gradually developed resistance to antibacterial drugs, especially the emergence of multidrug-resistant bacteria, which has brought severe challenges to clinical anti-infection treatment and hospital infection prevention and control. Therefore, it is very important to develop more accurate drug-resistant bacteria detection technology and drug-resistant bacteria sterilization technology that are closely related to food safety, disease prevention and clinical diagnosis and treatment. In the process of developing accurate detection and sterilization techno...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08J5/18C08J9/28C08J7/12C08J9/36C08L83/04C12N1/02C12R1/19
CPCC08J5/18C08J7/12C08J9/28C08J9/36C08J2383/04C12N1/02
Inventor 徐顺清沈先涛陈志亮杨柳倩
Owner HUAZHONG UNIV OF SCI & TECH
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