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Method for preparing electrochemical immunosensors for detecting food-borne pathogens on basis of quick scanning anodic stripping voltammetry technologies and application of electrochemical immunosensors

A technology of food-borne pathogenic bacteria and anode stripping voltammetry, which is applied in the field of preparation of electrochemical immunosensors, can solve the problems of undisclosed electrochemical immunosensors, complicated detection steps and high false positive probability, and achieves reliable detection results, Detect fast, highly sensitive effects

Inactive Publication Date: 2018-08-07
NINGBO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Common food-borne pathogens include: Vibrio parahaemolyticus, Vibrio vulnificus, Staphylococcus aureus, Escherichia coli, Salmonella, etc. The traditional method for detecting food-borne pathogens is the biochemical culture identification method. Complicated, long detection cycle, time-consuming and labor-intensive
With the rapid development of molecular biology techniques, methods such as polymerase chain reaction (PCR), DNA hybridization and loop-mediated isothermal amplification (LAMP), and biochips have also been applied to the detection of foodborne pathogens, and relatively good results have been obtained. Good accuracy and sensitivity, but there are many problems in practical applications: high false positive probability, expensive equipment, high detection cost, complicated detection steps, long detection time, etc.
At present, there are no relevant research reports on the preparation method of electrochemical immunosensors based on fast scanning anodic stripping voltammetry to detect food-borne pathogens at home and abroad.

Method used

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  • Method for preparing electrochemical immunosensors for detecting food-borne pathogens on basis of quick scanning anodic stripping voltammetry technologies and application of electrochemical immunosensors
  • Method for preparing electrochemical immunosensors for detecting food-borne pathogens on basis of quick scanning anodic stripping voltammetry technologies and application of electrochemical immunosensors
  • Method for preparing electrochemical immunosensors for detecting food-borne pathogens on basis of quick scanning anodic stripping voltammetry technologies and application of electrochemical immunosensors

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Experimental program
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specific Embodiment 1

[0037] A method for preparing an electrochemical immunosensor for detecting food-borne pathogenic bacteria based on a fast-scanning anode stripping voltammetry technique, comprising the following steps:

[0038] (1) Capture unit (cAb@Fe 3 o 4 ) preparation (preparation process such as figure 1 shown)

[0039] a. Add 0.1~0.3 g FeCl 2 • 4H 2 O and 0.6-0.9 g FeCl 3 •6H 2 O was dissolved in 40-50 mL of deoxygenated secondary water, stirred under the protection of nitrogen to mix evenly, and 28 wt% ammonia solution was added dropwise until the pH of the reaction solution was 10, and the temperature was raised to 80 °C and maintained at this temperature for 2 h. , after the reaction, cooled to room temperature under the protection of nitrogen, washed with water until neutral, and set the volume to 50 mL to obtain Fe 3 o 4 Nanoparticle solution;

[0040] b. Add 40-50 mL Fe 3 o 4 Nanoparticle solution was mixed with 0.4-0.6 mL 3-aminopropyltriethoxysilane (APTES), ultrasoni...

specific Embodiment 2

[0051] The method for detecting food-borne pathogenic bacteria using the electrochemical immunosensor prepared in specific example 1, the principle is as follows image 3 shown, including the following steps

[0052] The electrochemical immunosensor prepared in the above-mentioned specific example 1 is used as the working electrode, the platinum wire electrode is used as the counter electrode, and the 0.1M KCl solution is used as the supporting electrolyte solution. The fast-scanning anode stripping voltammetry analysis is carried out at a voltage scanning speed of 100 V / s. , measuring the anodic stripping peak current i p (μA); Determination of the anodic dissolution peak current corresponding to a series of different concentrations of food-borne pathogenic bacteria i p the size of the build ip The quantitative relationship between foodborne pathogenic bacteria concentration (cfu / mL); according to the quantitative relationship, the concentration of foodborne pathogenic ba...

specific Embodiment 3

[0054] The preparation and detection application of an electrochemical immunosensor for detecting Vibrio vulnificus based on a fast-scanning anode stripping voltammetry technique specifically includes the following steps:

[0055] (1) Capture unit (cAb@Fe 3 o 4 ) preparation (preparation process such as figure 1 shown)

[0056] a. Add 0.2 g FeCl 2 • 4H 2 O and 0.8 g FeCl 3 •6H 2 O was dissolved in 45 mL of deoxygenated secondary water, stirred under the protection of nitrogen to mix evenly, and 28 wt% ammonia solution was added dropwise until the pH of the reaction solution was 10, the temperature was raised to 80 °C and maintained at this temperature for 2 h, and the reaction was completed. Afterwards, cool to room temperature under the protection of nitrogen, wash with water until neutral, and set the volume to 50 mL to obtain Fe 3 o 4 Nanoparticle solution;

[0057] b. Add 45 mL of Fe 3 o 4 The nanoparticle solution was mixed with 0.5 mL 3-aminopropyltriethoxysil...

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Abstract

The invention discloses a method for preparing electrochemical immunosensors for detecting food-borne pathogens on the basis of quick scanning anodic stripping voltammetry technologies and applicationof the electrochemical immunosensors. The method is characterized by comprising steps of (1), adding Fe3O4 amide nano-particle solution into glutaraldehyde solution, carrying out reaction, magnetically separating and cleaning first reaction products, then adding food-borne pathogen capture antibodies into the first reaction products, carrying out reaction, and magnetically separating and cleaningsecond reaction products to obtain capture unit solution; (2), binding a composite nano-material AgNPs@g-C3N4 with food-borne pathogen antibodies to obtain signal units; (3), sequentially dropwise adding capture units, food-borne pathogen sample solution and the signal units to the surfaces of magnetic glassy carbon electrodes to obtain the electrochemical immunosensors. The application of the electrochemical immunosensors includes measuring the concentration of the food-borne pathogens in unknown samples according to quantitative relationships between anodic stripping peak currents ip and the concentration of the food-borne pathogens. The method and the application have the advantages of high sensitivity, specificity and detection speeds, reliable detection results and simple steps.

Description

technical field [0001] The invention relates to a detection method of food-borne pathogenic bacteria, in particular to a preparation method and application of an electrochemical immunosensor for detecting food-borne pathogenic bacteria based on a rapid scanning anode stripping voltammetry technique. Background technique [0002] Foodborne pathogens refer to pathogenic bacteria that can cause food poisoning or use food as a medium of transmission. Food is easily contaminated by food-borne pathogenic bacteria in the process of collection, processing, transportation, etc., resulting in frequent food poisoning and disease outbreaks, and the annual economic losses caused by food-borne pathogenic bacteria in my country are as high as $17 billion. Common food-borne pathogens include: Vibrio parahaemolyticus, Vibrio vulnificus, Staphylococcus aureus, Escherichia coli, Salmonella, etc. The traditional method for detecting food-borne pathogens is the biochemical culture identification...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/48G01N27/327
CPCG01N27/48G01N27/3275G01N27/3277
Inventor 宋信信郭智勇孟维强杨帆胡宇芳王邃
Owner NINGBO UNIV
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