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Gold nanorod biotic compound and preparation method and applications thereof

A gold nanorod and biological technology, applied in the field of electrochemical analysis, can solve the problems of error, low sensitivity, interference false positive, etc., and achieve the effect of high accuracy, high sensitivity and strong specificity

Inactive Publication Date: 2015-04-08
JIANGSU UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although these methods have played a great role in the detection of E. coli, they have disadvantages such as long time-consuming, low sensitivity, and cumbersome operation. To a certain extent, they cannot meet the needs of dairy product manufacturers, supervision and management departments for rapid detection.
For example, MPN counts each sample needs to go through serial dilution, initial fermentation, plate separation and re-fermentation, and it usually takes 3–5 days to get the test results. Some dairy companies often cause product inventory accumulation, which greatly limits the product quality. time to market; moreover, there are many interfering factors that affect the accuracy of the experimental results, such as the technical level of the operator for isolating a single colony, the selection of a single colony, and Gram staining techniques, etc., and its technical characteristics determine that this method sometimes It will also underestimate the number of E. coli in the sample and even cause missed detection
Moreover, the composition of dairy products is very complex, and other microorganisms will interfere with the test results and cause false positive errors

Method used

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  • Gold nanorod biotic compound and preparation method and applications thereof
  • Gold nanorod biotic compound and preparation method and applications thereof
  • Gold nanorod biotic compound and preparation method and applications thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0021] Embodiment 1 preparation d Ab-AuNR-Fc biocomplex:

[0022] Step 1: First, hexadecyltrimethylammonium bromide-coated gold nanorods (AuNR-CTAB) were prepared according to the classic seed method. Second, the preparation of AuNRsSiO 2 : Centrifuge 10 mL of AuNR-CTAB at 9,000 rpm for 10 minutes, add 10 mL of ultrapure water to resuspend, repeat 2 times. Under stirring conditions, add 100 μL sodium hydroxide (0.10 mol / L) to 10 mL AuNR-CTAB aqueous solution, and then add 60 μL ethanol solution of ethyl orthosilicate / 3-aminopropyltriethoxysilane ( Volume ratio: 1:1:5), continue to stir for 24 hours and collect to obtain AuNRsSiO 2 , and then washed with ultrapure water and dispersed in 5.0 mL of PBS for use;

[0023] Step 2: Take 1.0 mL of AuNRsSiO 2 solution, under stirring conditions, add 1.0 mL EDC / NHS solution (molar ratio: 4:1) to react for 10 minutes, then add 10 μL d Ab (4.0 mg / mL), 1.0 mL Fc saturated aqueous solution, continue to stir for 2 hours to prepare d A...

Embodiment 2

[0025] Example 2 is based on d Ab-AuNR-Fc biocomplex to construct an electrochemical immunosensor for the detection of Escherichia coli in dairy products

[0026]The cysteine-modified gold electrode was used as the working electrode, and the carboxyl group at the end of the cysteine ​​was activated with 0.40 mol / L EDC-0.10 mol / L NHS, and then combined with the capture antibody to prepare the immunosensor. Based on the specific interaction between the antibody and E. coli, the immune binding of E. coli, and then adsorption d Ab-AuNR-Fc biocomplex constructs "sandwich" immunoassay mode. The modified gold electrode was used as the working electrode, the saturated calomel electrode was used as the reference electrode, and the platinum wire was used as the auxiliary electrode. The Fc on the electrode surface was measured by differential pulse voltammetry to obtain the current signal, and the scanning potential was 0-0.70 V. As the concentration of Escherichia coli in the solution...

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Abstract

The invention discloses a gold nanorod biotic compound and a preparation method and applications thereof and belongs to the technical field of electrochemical analysis. A gold nanorod modified by silicon dioxide (AuNRs@SiO2) is used as a carrier and combined with a detection antibody (dAb) and ferrocenecarboxylic acid (Fc) to prepare dAb-AuNR-Fc which is used for amplifying escherichia coli in an electrochemical immunoassay dairy product. A 'sandwich' immunoassay model is established on basis of specificity interaction between the escherichia coli and escherichia coli antibody, and Fc combined with an electrode surface is measured by a differential pulse voltammetry to obtain a current signal. According to the gold nanorod biotic compound and the preparation method and the applications thereof, the electrochemical immunoassay method based on dAb-AuNR-Fc biotic compound is used for escherichia coli detection, the detection sensitivity, the specificity and the accuracy are high, and a novel method is provided for analysis and research of the escherichia coli in the dairy product.

Description

technical field [0001] The invention relates to an electrochemical immunoassay method, specifically, to a research on the establishment of an electrochemical immunosensing technology using nanocomposite materials, which is applied to the detection of Escherichia coli in dairy products, and belongs to the technical field of electrochemical analysis. Background technique [0002] Dairy products are rich in nutrients such as protein and fat, and are easily contaminated by pathogenic bacteria during their production, processing, transportation and storage, which will affect people's quality of life and health. Various surveys have shown that diarrhea and large-scale food poisoning caused by humans through milk-derived routes are related to the number of E. coli in dairy products. Therefore, E. coli has become an important indicator for evaluating the hygienic quality of dairy products and is used by countries all over the world. Microbiological standards for dairy products are l...

Claims

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Application Information

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IPC IPC(8): B22F1/02G01N33/569G01N27/48B82Y30/00B82Y40/00
Inventor 鲁文杰张新爱申建忠蒋玉香
Owner JIANGSU UNIV
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