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Method for detecting escherichia coli O157: H7 based on core-shell gold-platinum nanocluster

A technology of Escherichia coli and O157, which is applied in the field of microbial detection technology and pretreatment, can solve problems such as high technical difficulty, and achieve the effects of simplified operation steps, high stability, and shortened detection time

Pending Publication Date: 2021-11-16
HEFEI UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the choice of which recognition molecule, mimic enzyme and the coupling method between them can not only ensure the easy synthesis of the material but also ensure the sensitivity and stability of the color reaction, and realize the rapid and sensitive detection of Escherichia coli O157:H7 , has high technical difficulty and has not been reported

Method used

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  • Method for detecting escherichia coli O157: H7 based on core-shell gold-platinum nanocluster
  • Method for detecting escherichia coli O157: H7 based on core-shell gold-platinum nanocluster
  • Method for detecting escherichia coli O157: H7 based on core-shell gold-platinum nanocluster

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Embodiment 1

[0046] Application of the present invention in detection of Escherichia coli O157:H7 content in milk

[0047] When the present invention is used to detect the content of Escherichia coli O157:H7 in milk, it is implemented through the following steps: sample pretreatment, detection by the method of the present invention, and result analysis.

[0048] (1) Sample pretreatment

[0049] Take 1 mL of the purchased aseptic milk sample, add it to 9 mL of sterile PBS, mix well, and then add different concentrations of bacteria to prepare a bacterial suspension for later use.

[0050] (2) Carry out the detection of Escherichia coli O157:H7 content in the above-mentioned sample with detection method of the present invention

[0051] (1) Coated ovalbumin

[0052] Dilute ovalbumin to 30 mg / mL with 0.01 mol / L phosphate buffer solution with a pH value of 9.0 as the coating solution on the microtiter plate, and coat overnight at 4°C, then discard the coating solution. Washing the microtite...

Embodiment 2

[0069] A method for detecting Escherichia coli O157:H7 based on nucleoshell-type gold-platinum nanoclusters, the method is aimed at the detection of Escherichia coli O157:H7 antigens, in which nucleic acid aptamers serve as recognition elements, and gold-platinum nanoclusters as peroxidases.

[0070] see figure 1 , the specific detection steps are as follows:

[0071] (1) Coated ovalbumin

[0072] Dilute ovalbumin to 30 mg / mL on a microtiter plate with phosphate buffer solution with a concentration of 0.01 mol / L and a pH value of 9.0 as the coating solution, and coat at 4°C overnight, discard the coating solution, and wash with Wash the plate with liquid. The amount of aptamer-modified magnetic beads was 100 μg / well, and the amount of aptamer-modified gold-platinum nanoclusters was 10 pmol / L.

[0073] (2) Adding aptamer-modified magnetic beads and aptamer-modified gold-platinum nanoclusters

[0074] Aptamer-modified magnetic beads and aptamer-modified gold-platinum nanoc...

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Abstract

The invention relates to a method for detecting escherichia coli O157: H7 based on a core-shell gold-platinum nanocluster, and belongs to the technical field of microbiological detection and pretreatment. According to the method, the core-shell type gold-platinum nanocluster is used as mimic enzyme, streptavidin is adsorbed on the surface of the gold-platinum nanocluster through electrostatic interaction, and then a biotinylated specific aptamer and the gold-platinum nanocluster are combined so as to serve as a probe, so that specific recognition of the escherichia coli O157: H7 is promoted. On the basis of a sandwich method strategy, an aptamer modified magnetic bead is used as a capture probe, interference of complex matrixes in a sample on a detection result is reduced through a magnetic attraction effect; a biotin-avidin system is adopted to modify the aptamer on the surface of the gold-platinum nanocluster, so that steric hindrance is reduced, and the loading capacity of recognition molecules is improved. According to the method of the invention, synchronous recognition of the escherichia coli O157: H7 is realized, the operation steps are simplified, the reaction time is greatly shortened, the detection sensitivity is improved to 102 CFU / mL, the cost is reduced, the influence of a sample matrix is reduced, and the detection efficiency is improved. The method has a bright application prospect.

Description

technical field [0001] The invention belongs to the field of microbial detection technology and pretreatment technology, and in particular relates to a detection method for Escherichia coli O157:H7 based on core-shell gold and platinum nanoclusters. Background technique [0002] Escherichia coli O157:H7 ( Escherichia coli O157:H7) belongs to the genus of Enterobacteriaceae Escherichia, Gram-negative bacilli, without spores, with flagella; resistant to acid and low temperature, among which serotype O157:H7 is the main pathogenic bacteria of enterohemorrhagic and enteritis; can It is transmitted through foods such as milk, chicken, beef, and fruit. The Vero toxin and Shiga-like toxin produced by it can cause symptoms such as hemorrhagic colitis, hemolytic uremic syndrome, and thrombotic thrombocytopenic purpura in infected persons, making them susceptible The crowd is mainly children and the elderly. The infection dose of this bacterium is extremely low, the infectivity is s...

Claims

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Application Information

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IPC IPC(8): G01N21/31G01N21/78
CPCG01N21/31G01N21/78Y02A50/30
Inventor 高宝叶应旺凌娜沈益忠张丹凤偶德馨王洋
Owner HEFEI UNIV OF TECH
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