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Preparation method and detection method of nano immunosensor used for rapid detection of enterobacter sakazakii

A technology of Enterobacter sakazakii and immunosensor, which is applied in the field of nano-immunosensor preparation, can solve the problems of difficult to distinguish results, cumbersome operation, and long time consumption, and achieve the effects of improving detection efficiency, simple preparation, and shortened inspection time

Active Publication Date: 2015-11-18
HEBEI SANYUAN FOOD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the traditional detection method has disadvantages such as cumbersome operation, long time-consuming, and difficult to distinguish the results.

Method used

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  • Preparation method and detection method of nano immunosensor used for rapid detection of enterobacter sakazakii
  • Preparation method and detection method of nano immunosensor used for rapid detection of enterobacter sakazakii
  • Preparation method and detection method of nano immunosensor used for rapid detection of enterobacter sakazakii

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Detection of Enterobacter sakazakii in milk powder sample I by immunosensing

[0029] 1. Preparation of Immunosensors

[0030] a. Electrode pretreatment

[0031] Polish the glassy carbon electrode with alumina polishing powder to the mirror surface, wash it with distilled water, then use secondary deionized water, absolute ethanol, and secondary deionized water to ultrasonically clean it for 3 minutes to 5 minutes, and dry the glassy carbon electrode with nitrogen . Then, put the phosphate buffer solution with a pH value of 7.0 into the beaker, then add potassium ferricyanide, potassium chloride and hydrogen peroxide to make it 2mmol / L, 0.1mol / L and 0.5mmol / L respectively, Get the electrolyte solution, then insert the reference electrode, the counter electrode and the glassy carbon electrode into the electrolytic cell of the electrolyte solution, connect the PAR270 electrochemical workstation; use the PAR270 electrochemical workstation to perform cyclic voltammetry sc...

Embodiment 2

[0042] Detection of Enterobacter sakazakii in milk powder sample II by immunosensing

[0043] 1. Preparation of Immunosensors

[0044] a. Electrode pretreatment

[0045] Same as Example 1

[0046] b. Preparation of graphene-polyamide-amine composite

[0047] Same as Example 1

[0048] c. Preparation of Immunosensors

[0049] Use a micropipette gun to accurately measure 4 μL of the graphene-polyamide-amine nanocomposite obtained in step b and drop-coat it on the surface of the glassy carbon electrode obtained in step a, dry it naturally or in an infrared dryer at room temperature, and then The surface of the glassy carbon electrode was drop-coated with horseradish peroxidase-labeled Enterobacter sakazakii antibody in the same volume as the graphene-polyamide-amine nanocomposite, and placed in a refrigerator at 4°C for 1.5h to 2h, and then drop-coated The same volume of 1wt% bovine serum albumin solution was treated in a 37°C incubator for 55min to 60min to block the non-sp...

Embodiment 3

[0055] Detection of Enterobacter sakazakii in milk powder sample III by immunosensing

[0056] 1. Preparation of Immunosensors

[0057] a. Electrode pretreatment

[0058] Same as Example 1

[0059] b. Preparation of graphene-polyamide-amine composite

[0060] Same as Example 1

[0061] c. Preparation of Immunosensors

[0062] Use a micropipette to accurately measure 7 μL of the graphene-polyamide-amine nanocomposite obtained in step b and drop-coat it on the surface of the glassy carbon electrode obtained in step a, dry it naturally or in an infrared dryer at room temperature, and then The surface of the glassy carbon electrode was drop-coated with horseradish peroxidase-labeled Enterobacter sakazakii antibody in the same volume as the graphene-polyamide-amine nanocomposite, and placed in a refrigerator at 4°C for 1.5h to 2h, and then drop-coated The same volume of 1wt% bovine serum albumin solution was treated in a 37°C incubator for 55min to 60min to block the non-speci...

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Abstract

The invention belongs to the technical field of food safety rapid detection, and more specifically relates to a preparation method and a detection method of a nano immunosensor used for rapid detection of enterobacter sakazakii. According to the preparation method, a glassy carbon electrode is taken as a base electrode; a polyamide dendritic macromolecule material combined reductive graphene nano composite material is taken as an electroactivity modification membrane layer, and is immobilized onto the surface of the glassy carbon electrode; physical absorption or bovine serum albumin-glutaraldehyde crosslinking is adopted to immobilize horse radish peroxidase-labelled enterobacter sakazakii antibody so as to obtain the nano immunosensor. According to the detection method, the nano immunosensor is applied to enterobacter sakazakii detection in food, and obtained results accord with results obtained via a national standard method; the detection method is high in sensitivity, specificity, and accuracy, is short in detection time, is simple, convenient, and rapid, and possesses a promising application prospect.

Description

technical field [0001] The invention relates to a preparation method of a nano-immunosensor for rapid detection of Enterobacter sakazakii and a method for detecting Enterobacter sakazakii in milk powder by using the nano-immune sensor. Background technique [0002] Enterobacter sakazakii is a common Gram-negative abacillus that parasitizes in the intestines of humans and animals. It has perinatal flagella, is motile, and is facultatively anaerobic. It is one of the normal intestinal flora. Enterobacter sakazakii, as an opportunistic pathogen, can cause severe meningitis, enterocolitis, and bacteremia in neonatal children. And can lead to neurological sequelae and even death, the mortality rate is as high as more than 50%. Infant milk powder, as the main type of food carrying Enterobacter sakazakii, has caused a number of incidents related to food safety in recent years, and has attracted great attention from relevant international organizations, governments and consumers. ...

Claims

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Application Information

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IPC IPC(8): G01N27/48G01N33/569
Inventor 李书国李艳娟李朝旭房新平张媛媛
Owner HEBEI SANYUAN FOOD
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