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Method for detecting staphylococcus aureus

A staphylococcus, golden yellow technology, applied in the field of microbial detection, can solve the problems of high sensitivity requirements, long detection time, inability to distinguish the activity of pathogenic bacteria, etc., and achieve the effect of good stability and high sensitivity

Inactive Publication Date: 2012-08-22
沈鹤柏
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] The technical problem to be solved by the present invention is to provide a method for detecting Staphylococcus aureus and its special detection test paper to overcome the high sensitivity requirements, complicated enrichment process, long detection time and inability to distinguish pathogenic bacteria in the existing detection methods. Bacterial activity, and the need for large-scale equipment and other defects

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  • Method for detecting staphylococcus aureus
  • Method for detecting staphylococcus aureus
  • Method for detecting staphylococcus aureus

Examples

Experimental program
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Effect test

Embodiment 1

[0046] Preparation of Immunized Superparamagnetic Nanomagnetic Beads and Enrichment of Bacteria

[0047] 1. Magnetic core Fe 3 o 4 Synthesis and silicon-coated

[0048] Separately prepare FeSO with double distilled water 4 ·7H 2 O, FeCl 3 ·6H 2 O, 2.5mol / L NaOH three kinds of solutions, the FeSO 4 ·7H 2 O, FeCl 3 ·6H 2 O The two solutions are mixed so that the concentration of ferrous ions in the mixed solution is 0.15mol / L, and the concentration of ferric ions is 0.25mol / L, and the NaOH solution configured above is slowly added dropwise under vigorous stirring into the mixed iron salt solution, age the obtained precipitate at 60°C for 2 hours, wash the precipitate with double distilled water for 3 to 5 times, filter and dry at 60°C for 24 hours, and grind it in an agate mortar to obtain the product. Dry at 60°C and store at room temperature for later use.

[0049] Due to the magnetic core Fe 3 o 4 The dispersibility is poor, the particle size is uneven, and it is...

Embodiment 2

[0054] Preparation of Colloidal Gold Immunochromatographic Test Paper for Staphylococcus aureus

[0055] 1. Preparation of colloidal gold-labeled Staphylococcus aureus monoclonal antibody complex

[0056] Label the Staphylococcus aureus monoclonal antibody on the prepared colloidal gold surface with an average particle size of 10-80nm, then wash twice with 0.002M boric acid buffer solution, and concentrate to 1 / 50-1 / 2. Generally 1 / 25, store at 4°C for later use.

[0057] 2. Preparation of colloidal gold immunochromatography test paper

[0058]2.1 Preparation of sample pad: In this embodiment, glass cellulose film is selected as the sample pad material, and the glass cellulose film is put into the sample pad treatment solution (0.01M PBS (PH=7.4), containing 1% BSA, 0.1% Trion Soak in X-100) and dry at 30-45°C, usually 37°C.

[0059] 2.2 Preparation of conjugation pad: In this example, glass cellulose membrane was selected as the sample pad material, and the colloidal gold-...

Embodiment 3

[0064] 1, carry out specificity test to test paper of the present invention by detecting 90 parts of food samples

[0065] 50 food samples containing Staphylococcus aureus, 10 food samples containing Escherichia coli O157:H7, 10 food samples containing Listeria monocytogenes, 10 food samples of Enterobacter sakazakii, and 10 blank cultures As the sample to be tested, 5ml was taken out and mixed with 50μl superparamagnetic nanoparticles coated with Staphylococcus aureus specific antibody after ultrasonication for 10min respectively to form a complex of antibody-magnetic particle-Staphylococcus aureus; Under the action of attraction, the complex is collected; the complex is inoculated into 7.5% sodium chloride broth culture medium for 6-12 hours, and the antibody-magnetic particle-Staphylococcus aureus is magnetically separated under a 3000Gs magnetic field The complex, the bacterial solution after magnetic separation was centrifuged at 15000r / min for 20min, then dissolved in 0....

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Abstract

The invention relates to a method for detecting staphylococcus aureus, namely a method for enriching and screening the staphylococcus aureus by a one-step method. The method comprises the following steps of: utilizing a superparamagnetic nano particle covered by a staphylococcus aureus specific antibody to be specially combined with the staphylococcus aureus to form a composite; collecting a superparamagnetic nano particle composite under the attraction effect of an external magnetic field; inoculating the enriched composite into a culture medium to culture for 6-12 hours; and utilizing a staphylococcus aureu colloidal gold colloidal gold test paper to carry out qualitative and quantitative detection. The invention further provides the corresponding colloidal gold colloidal gold test paper for rapidly detecting the staphylococcus aureus, which has the flexibility of 104 cfu / mL of bacteria liquid and can be used for food safe detection, clinical rapid diagnosis, and onsite rapid screening of samples including water quality and the like.

Description

technical field [0001] The invention relates to the field of microbial detection, in particular to a method for rapid enrichment, screening, qualitative and quantitative detection of Staphylococcus aureus and special test paper. Background technique [0002] People depend on food, and food is the most basic material condition for the survival and development of human beings. However, food safety accidents and public health emergencies caused by food-borne pathogenic microorganisms in the world and in my country have caused people to Highly concerned about food safety, according to the statistics of the United Nations World Health Organization, the under-reporting rate of food-borne diseases is over 95%, and the infection and food poisoning caused by food-borne pathogenic microorganisms exceed 85%. The rapid detection of food-borne pathogens has always been the focus of research. The rapid detection of food-borne pathogens is the use of microbiological, chemical, biochemical, ...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/569G01N33/558
Inventor 牛凯莉支援徐宽沈鹤柏
Owner 沈鹤柏
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