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A method for specific detection of Staphylococcus aureus

A staphylococcus, golden yellow technology, applied in the direction of measuring devices, DNA/RNA fragments, instruments, etc., can solve the problems of indistinguishable strain types, etc., and achieve the effects of simplifying sample pretreatment, improving sensitivity, and specific detection

Active Publication Date: 2017-06-13
JIANGNAN UNIV
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  • Abstract
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  • Application Information

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Problems solved by technology

Although the ATP bioluminescence method has the characteristics of rapid detection, it cannot distinguish the strain type

Method used

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  • A method for specific detection of Staphylococcus aureus
  • A method for specific detection of Staphylococcus aureus
  • A method for specific detection of Staphylococcus aureus

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Embodiment Construction

[0024] The present invention is further described in conjunction with embodiment:

[0025] (1) Synthesis of Fe by solvothermal method 3 o 4 magnetic nanoparticles and modify their surfaces. Analytical balance accurately weighs 0.81g FeCl 3 ·6H 2 O, 2.16g of NaAC was added to 30mL of ethylene glycol, stirred magnetically at 50°C to form a homogeneous solution. React with the reactor at 200°C for 6h. After natural cooling, cross-wash with water and ethanol three times each, and vacuum-dry at 50°C for 12 hours to obtain Fe 3 o 4 magnetic nanoparticles.

[0026] (2) Using the improved Stober method for Fe 3 o 4 The surface of magnetic nanoparticles is modified by amination. Take 30mg of Fe synthesized above 3 o 4 Magnetic nanoparticles, add 100mL, 0.1M HCl, ultrasonic dispersion 5min. Wash twice with water and once with ethanol. Disperse in 126.6mL absolute ethanol and 30mL ultrapure water, stir mechanically (30 rpm), add 1.05mL concentrated ammonia water, and then s...

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Abstract

The invention provides a method for specific detection of staphylococcus aureus. The method is characterized in that a nucleic acid aptamer is modified on the surfaces of magnetic nano particles, quickly recognizes, and is combined with staphylococcus aureus in a solution sample; target fungi combined with the magnetic nano particles are separated and enriched by using a magnetic field; captured fungi are suspended in a lysate with a certain volume; ATP (Adenosine Triphosphate) extracting staphylococcus aureus is released and used; and as the amount of ATP is directly proportional to the concentration of fungi liquid, after a linearity curve of the fungi concentration and the luminous intensity is obtained by an ATP bioluminescence method, staphylococcus aureus can be detected quantitatively. The method has the advantages that the aptamer is easy to synthesize, modify and fix, and can be kept for a long time; by performing combination, separation, enrichment and concentration on the target fungi, sample matrices and other interference fungi are removed effectively; and the detection sensitivity is improved.

Description

[0001] Technical field: [0002] The invention belongs to the field of food safety, and in particular relates to a detection method for Staphylococcus aureus in food. [0003] Background technique: [0004] Staphylococcus aureus is a Gram-positive bacterium that causes purulent infections in humans and animals. It is widely distributed in nature, such as air, soil, water, and human and animal excrement. The bacterium is not strict to the growth environment and is an conditional pathogenic bacterium. Most food substrates such as milk, meat, eggs, fish and other products provide good conditions for the growth of Staphylococcus aureus. Toxins can cause food poisoning, accompanied by severe vomiting, dehydration and collapse. According to statistics, food poisoning caused by Staphylococcus aureus ranks in the top three of bacterial food poisoning in all countries in the world, accounting for about 25%, and causes huge economic losses, seriously affecting human food safety and econ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/115
CPCG01N33/54326G01N33/54346G01N33/56938G01N2333/31
Inventor 沈晓芳陈文玲庞月红
Owner JIANGNAN UNIV
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