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Method for rapidly detecting pathogenic bacteria

A technology for pathogenic bacteria and food-borne pathogenic bacteria, applied in the field of microbial detection, can solve the problem of high dosage of nanoparticles

Inactive Publication Date: 2015-07-01
SHANGHAI INST OF MICROSYSTEM & INFORMATION TECH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method is sensitive, rapid and specific. However, the preparation method of this nanoparticle is limited to the detection of three pathogenic bacteria, and the amount of nanoparticle is very high (particles / bacteria = 10000 / 1)

Method used

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  • Method for rapidly detecting pathogenic bacteria
  • Method for rapidly detecting pathogenic bacteria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1: Preparation of four bifunctional silicon-based nanospheres with different optical properties

[0034] In the present invention, the bifunctional nanosphere is improved by obtained by the method. Specifically, it is obtained by hydrolyzing ethyl orthosilicate in ethanol solution under the catalysis of a certain concentration of ammonia water. By adjusting the relative concentrations of ammonia water, tetraethyl orthosilicate, ethanol, water, magnetic nanoparticles and quantum dots, the bifunctional nanospheres meeting the detection requirements are obtained. Add quantum dots with different optical properties (such as emission wavelengths of 481nm, 518nm, 565nm and 610nm), thereby changing the optical properties of the bifunctional composite nanospheres, and obtaining four bifunctional nanospheres with different optical properties (marked as nanospheres 481 , nanosphere 518 , nanosphere 565 , nanosphere 610 ). A typical synthesis process is as follows: ...

Embodiment 2

[0036] Example 2: Four kinds of immune bifunctional nanospheres capturing enterohaemorrhagic Escherichia coli (E.coli O157:H), Salmonellae, Listeria and Campylobacter respectively Probe construction

[0037] Firstly, the surface of the above four kinds of bifunctional composite nanospheres was modified. Specifically, 0.5 g of nanospheres were dissolved in 50 mL of pyridine, 0.2 g of succinic anhydride and 0.02 g of 4-dimethylaminopyridine were added, stirred at room temperature overnight, magnetically separated, and methanol Wash twice with water.

[0038] Then, enterohemorrhagic Escherichia coli antibody, Salmonella antibody, Listeria monocytogenes antibody and Campylobacter antibody were covalently linked to the above four bifunctional nanospheres to obtain four kinds of immune bifunctional nanosphere probes (labeled for nanospheres 481 -E, nanospheres 518 -S, nanospheres 565 -L, nanosphere 610 -C). The operation steps are as follows:

[0039] Take 500μg of nanosphere...

Embodiment 3

[0040] Example 3: Four immune quantum dot probes for respectively labeling E.coli O157:H, Salmonellae, Listeria and Campylobacter build

[0041] The monoclonal antibodies that specifically recognize the target foodborne pathogenic bacteria enterohemorrhagic Escherichia coli antibody, Salmonella antibody, Listeria monocytogenes antibody and Campylobacter antibody were connected with the corresponding quantum dots to form four kinds of immune quantum Point probes (labeled as quantum dots 481 -E, quantum dots 518 -S, quantum dots 565 -L, quantum dots 610 -C). The size of the antibody and quantum dots labeled by this probe corresponds to the size of the labeled antibody and quantum dots embedded in the immune complex nanospheres, which can be similar to the formation of immune complex nanosphere probes, and the formation of target food-borne pathogenic bacteria Similar to a sandwich structure. The operation steps are as follows:

[0042] Take 500 μL of quantum dots in a 1.5...

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Abstract

The invention provides a method for rapidly detecting pathogenic bacteria. The method is characterized in that firstly, a difunctional composite nanosphere is prepared; quantum dots with different sizes and the composite functional nanosphere are connected with a monoclonal antibody capable of specifically recognizing target bacteria, so as to obtain immune quantum dot probes capable of carrying out antigen-antibody reaction with antigens on the surface of the target bacteria, an immune composite nanosphere probe, and other probes The composite nanosphere with the composite structure can be used as a carrier for immunologically recognizing and separating the pathogenic bacteria and also can be used as a signal enhancer element of the immune quantum dot probe, secondary amplification of a detection signal is realized, and the optical strength can be detected under the corresponding wavelength and multiple target microorganisms can be detected. According to the method, the detection time less than or equal to 2h can be greatly shortened, the sensitivity is improved and multiple pathogenic bacteria can be detected simultaneously through immunoseparation of the magnetic nanoparticles and the signal amplification effect of the quantum dots having dual labels, and the method is suitable for on-site rapid detection of foods and environment samples and can be popularized and applied in grass roots.

Description

technical field [0001] The present invention relates to a method for rapid detection of various pathogenic bacteria, more specifically, to a preparation of adjustable bifunctional nanospheres and its application in the rapid detection of various food-borne pathogenic bacteria, that is, the described The bifunctional composite nanosphere can be used in medical diagnosis, food safety, environmental monitoring and so on. It belongs to the field of microbiological detection. Background technique [0002] Bacterial pathogens seriously endanger human health. There are many kinds of foodborne pathogenic microorganisms, and the lack of sensitive, convenient and specific rapid detection technology is one of the main reasons why food safety cannot be effectively guaranteed. Therefore, it is an urgent need for food safety and national security to develop fast, sensitive and reliable detection methods and on-site and portable detection instruments for pathogenic bacteria. [0003] Th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N21/31B82Y15/00
CPCG01N33/577B82Y15/00G01N21/3103G01N2021/3129
Inventor 葛玉卿金庆辉毛红菊赵建龙
Owner SHANGHAI INST OF MICROSYSTEM & INFORMATION TECH CHINESE ACAD OF SCI
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