Nanoprobe for visual detection of staphylococcus aureus and preparation method thereof

A staphylococcus and nano-probe technology, applied in biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problem of less research and achieve the effect of wide detection limit

Inactive Publication Date: 2017-08-18
SICHUAN AGRI UNIV
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  • Abstract
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  • Claims
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Problems solved by technology

[0005] However, there are still very few studies on the visualization of gold nanoprobes for the detection of highly harmful Staphylococcus aureus.

Method used

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  • Nanoprobe for visual detection of staphylococcus aureus and preparation method thereof

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Embodiment 1

[0023] Probe preparation:

[0024] (1) Prepare gold nanoparticles with citric acid and tannic acid, and the particle size of the obtained gold nanoparticles is 20~30nm; then place the obtained gold nanoparticles in dihydrate bis(p-sulfonylphenyl)phenylphosphine Reaction in dipotassium salt solution;

[0025] (2) Mix the following three kinds of DNA with the result of step (1):

[0026] 5’-SH-TTT TTC TAC CGG TAC ATG GAC AGT ATG TCT TTC CAT ACT TGC ATG TCA CAGCGC-3’;

[0027] 5’-SH-TTT ATC TAC AGG CAC ATG TAC AGT ATG TCT TTC CAT ACT GTA AGT TGA CAGCGG-3’;

[0028] 5’-SH-TTT CTC TAC AGG CGC ATG TAC GGA ATG TCT TTC CAT ACT TTC AGT GAA CAGCCC-3’

[0029] After mixing, incubate at 37°C, then add SDS, add NaCl solution with a concentration of 0.3mol / L after shaking, and age at 37°C, remove the supernatant after centrifugation to remove excess DNA, and resuspend the pellet with PBS.

[0030] The concentration of the dipotassium salt solution of bis(p-sulfonylphenyl)phenylphosphine...

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Abstract

The invention belongs to the field of bacterium detection materials and particularly relates to a nanoprobe for visual detection of staphylococcus aureus and a preparation method thereof. The method comprises the following steps: (1) utilizing citric acid and tannic acid to prepare gold nanoparticles with the particle size of 20-30nm; and putting the obtained gold nanoparticles into a dihydrated bis(p-sulfonylphenyl)phenyl dipotassium phosphide solution for reaction; (2) mixing three special DNAs with a product obtained in the step (1), then carrying out incubation at the temperature of 37 DEG C, then adding SDS (Sodium Dodecyl Sulfonate), adding an NaCl solution with the concentration of 0.3-0.4mol / L after oscillation, carrying out aging treatment at the temperature of 37 DEG C, and after centrifugal treatment, removing the supernatant and redundant DNAs, and resuspending precipitates with PBS (Polybutylene Succinate). The nanoprobe and the preparation method have the main advantages that the detection limit is wider, and the lowest detection limit can reach 1*10-14mol / L(3S / N).

Description

technical field [0001] The invention belongs to the field of bacteria detection materials, and in particular relates to a nanometer probe for visual staphylococcus aureus detection and a preparation method thereof. Background technique [0002] At present, the detection of bacteria is usually carried out by PCR, which is time-consuming and labor-intensive. Since Staphylococcus aureus is a relatively strong pathogen, it is essential to detect it in food or in some special living and working environments. [0003] In recent years, visual inspection has been increasingly favored by researchers due to its convenience. [0004] Nanogold has many uses, including the detection of microorganisms. For example, Lu Weiping and others labeled monomaleic imide-modified nanogold on the nucleic acid molecules of pathogenic bacteria, and achieved signal amplification through gold-labeled silver staining technology. , to achieve the detection of some pathogenic bacteria. [0005] However,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/14
CPCC12Q1/6811C12Q1/689C12Q2563/137C12Q2563/155
Inventor 黎杉珊罗擎英刘耀文申光辉张志清吴贺君陈安均刘兴艳
Owner SICHUAN AGRI UNIV
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