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Three-dimensional cell paper chip sensor and application thereof in bacterial lipopolysaccharide detection

A three-dimensional cell and paper chip technology, applied in the field of analysis and detection, can solve the problems of repeated use, expensive electrodes, and influence on the accuracy of detection results, and achieve the effect of high sensitivity and low cost

Active Publication Date: 2019-12-31
南京市食品药品监督检验院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Commercial conventional electrodes are expensive and need to be used repeatedly after modification
Electrode cleaning and activation is complicated and time-consuming, if the regeneration is not good, it will affect the accuracy of the detection results

Method used

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  • Three-dimensional cell paper chip sensor and application thereof in bacterial lipopolysaccharide detection
  • Three-dimensional cell paper chip sensor and application thereof in bacterial lipopolysaccharide detection
  • Three-dimensional cell paper chip sensor and application thereof in bacterial lipopolysaccharide detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1: Construction of three-dimensional cell paper chip sensor

[0050] The construction process of the three-dimensional cell paper chip sensor is as follows: figure 2 As shown, the paper chip analysis device is prepared first, the paper chip is turned over, the pyrrole / graphene oxide dispersion is added to the electrode, the paper chip is turned over, the polypyrrole / graphene oxide is electropolymerized, Nafion is added to modify the working electrode, and the working electrode is turned over Paper chip, load the 3D cell culture system to the working electrode, turn over the paper chip, combine the paper chip with the conductive jacket, connect the electrochemical workstation, and construct the 3D cell paper chip sensor.

[0051] 1. Preparation of paper chip analysis device

[0052] Use CorelDRAW software to design hydrophilic / hydrophobic regions and electrode patterns; use a wax printer to print hydrophobic regions on double-circle filter paper, place them in ...

Embodiment 2

[0059] Example 2: Characterization of three-dimensional cell paper chip sensor

[0060]Electrochemical characterization of the constructed three-dimensional cell paper chip sensor. The test conditions of cyclic voltammetry are: the voltage scanning range is -0.2V to 0.6V, the scanning rate is 100mV / s, and the sampling interval is 1mV. The test conditions of differential pulse voltammetry are: the voltage sweep range is -0.2V to 0.6V, the pulse amplitude is 50mV, the pulse width is 50ms, the pulse period is 500ms, and the potential increment is 4mV. Fe(CN) 6 3- / 4- The electrolyte concentration was 2.5mmol / L, and all tests were performed at room temperature.

[0061] exist image 3 In A, curve a is the bare electrode in Fe(CN) 6 3- / 4- The reversible redox peak was obtained by scanning in the solution, and the peak current at this time was 11.12 μA (by image 3 B is calculated from the curve a in the DPV graph); curve b is the reversible redox peak of the polypyrrole / graph...

Embodiment 3

[0063] Example 3: Application of three-dimensional cell paper chip sensor:

[0064] 1. Detection of lipopolysaccharide

[0065] A standard lipopolysaccharide (L7770Sigma-Aldrich) derived from Salmonella Enteritidis was dissolved in PBS to prepare a 1 mg / mL stock solution. Diluted with DMEM medium to a concentration of 10 4 ng / mL, 10 3 ng / mL, 10 2 ng / mL, 50ng / mL, 10ng / mL, 4ng / mL, 3ng / mL, 2ng / mL, 1ng / mL, 0.1ng / mL, 0.01ng / mL lipopolysaccharide solution. Add the lipopolysaccharide solution dropwise to the working area of ​​the three-dimensional cell paper chip sensor to stimulate the cells to be detected.

[0066] 2. Analysis conditions and methods

[0067] Electrochemical detection parameters: Differential pulse voltammetry is adopted, the voltage scanning range is -0.2V~0.5V, the pulse amplitude is 50mV, the pulse width is 50ms, the pulse period is 500ms, the potential increment is 4mV, and the reaction medium is PBS Solution (135mM NaCl, 4.7mM KCl, 10mM NaCl 2 HPO 4 , 2...

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Abstract

The invention discloses a three-dimensional cell paper chip sensor and an application thereof in bacterial lipopolysaccharide detection, and belongs to the technical field of analysis and detection. The novel three-dimensional cell paper chip sensor is constructed by combining a wax spraying printing technology, a silk-screen printing technology, a cell three-dimensional culture technology and anelectrochemical sensing technology, has the advantages of low cost and portability of a paper chip, authenticity of three-dimensional cell culture and high sensitivity and rapidity of electrochemicalanalysis at the same time, and can accurately detect low-content lipopolysaccharide. The invention further discloses a preparation method and an application of the three-dimensional cell paper chip sensor, and indirect detection and toxicity judgment of food-borne pathogenic bacteria can be achieved. The three-dimensional cell paper chip sensor has the advantages of sensitivity, high efficiency, miniaturization and the like, is low in price, and is suitable for detection of known pathogenic bacteria and toxicity judgment of unknown pathogenic bacteria.

Description

technical field [0001] The invention relates to a three-dimensional cell paper chip sensor and its application in the detection of bacterial lipopolysaccharide, belonging to the technical field of analysis and detection. Background technique [0002] Gram-negative bacteria, as the most common pathogenic bacteria, have become a serious threat to food safety and public health. Although high temperature and high pressure can kill bacteria, bacterial endotoxin (lipopolysaccharide, LPS) will not disappear or be destroyed with the death of bacteria. LPS is covalently linked from the inside to the outside by three parts: the hydrophobic lipid A, the core oligosaccharide and the hydrophilic O-antigen. Its toxic components are mainly lipid A. Lipopolysaccharides cause an inflammatory response that can lead to many infections or fatal diseases. Therefore, early warning and detection of LPS bioactivity are as important as the identification of the bacteria themselves. Not only that...

Claims

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Application Information

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IPC IPC(8): G01N27/48G01N27/30G01N33/53
CPCG01N27/48G01N27/30G01N33/5302
Inventor 蒋卉杨军万凯
Owner 南京市食品药品监督检验院
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