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Electrochemical detection method of Staphylococcus aureus

A staphylococcus, detection method technology, applied in the field of microbial detection, can solve the problems of complex detection process steps, expensive reagents, expensive instruments, etc., and achieve good detection stability, fast and accurate detection, simple and fast detection

Active Publication Date: 2017-03-15
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Microbiological methods are time-consuming and cumbersome to operate
The ELISA method takes a relatively short time, but its operation skills are strong, the control of key points has a great impact on the results, and the detection limit is as high as 10 6 CFU / mL
Polymerase chain reaction (PCR) is the most mature detection method for Staphylococcus aureus researched outside the national standard method, but the reagents required by the PCR method are expensive, and the entire detection process is complicated
Because the method is extremely sensitive, exogenous DNA, improper control of test conditions, primer design and selection of target sequences will affect the experimental results
These techniques are faster than traditional microbiological methods and simpler than PCR methods, but require expensive instruments and have high detection limits, generally 10 5 ~10 6 CFU / ml

Method used

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  • Electrochemical detection method of Staphylococcus aureus
  • Electrochemical detection method of Staphylococcus aureus
  • Electrochemical detection method of Staphylococcus aureus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Dissolve 1.25g of sucrose and 0.14g of concentrated sulfuric acid in 5.0g of water, add 1g of SBA-15 to the solution, dry the mixture in an oven at 100°C for 6h, then heat the oven to 160°C for 6h; 0.75g of sucrose, Dissolve 0.08g of concentrated sulfuric acid in 5.0g of water, immerse the above material into the mixture, dry it in an oven at 100°C for 6h, then heat the oven to 160°C for 6h; calcinate the obtained product at 900°C in a nitrogen atmosphere for 6h, The heating rate is 2.5°C / min, and finally the product is soaked in hydrofluoric acid, washed and dried. Finally, ordered mesoporous carbon material CMK-3 was obtained.

[0027] Immerse 300mg of CMK-3 nanomaterials into 15mL of 1.75mol / L ammonium persulfate solution and 15mL of 2mol / L sulfuric acid solution, stir in a water bath at 40°C for 24h, wash and dry to obtain carboxyl-functionalized mesoporous Carbon material O-CMK-3.

[0028] 5mg EDC and 6mg NHS were added to 30mL Tris-HCL buffer, 30mg carboxyl-func...

Embodiment 2

[0032] Dissolve 1.25g of sucrose and 0.14g of concentrated sulfuric acid in 5.0g of water, add 1g of SBA-15 to the solution, dry the mixture in an oven at 100°C for 6h, then heat the oven to 160°C for 6h; 0.75g of sucrose, Dissolve 0.08g of concentrated sulfuric acid in 5.0g of water, immerse the above material into the mixture, dry it in an oven at 100°C for 6h, then heat the oven to 160°C for 6h; calcinate the obtained product at 900°C in a nitrogen atmosphere for 6h, The heating rate is 2.5°C / min, and finally the product is soaked in hydrofluoric acid, washed and dried. Finally, ordered mesoporous carbon material CMK-3 was obtained.

[0033] Immerse 300mg of CMK-3 nanomaterials into 15mL of 1.75mol / L ammonium persulfate solution and 15mL of 2mol / L sulfuric acid solution, stir in a water bath at 40°C for 24h, wash and dry to obtain carboxyl-functionalized mesoporous Carbon material O-CMK-3.

[0034]5mg EDC and 6mg NHS were added to 30mL Tris-HCL buffer, 30mg carboxyl-funct...

Embodiment 3

[0038] Dissolve 1.25g of sucrose and 0.14g of concentrated sulfuric acid in 5.0g of water, add 1g of SBA-15 to the solution, dry the mixture in an oven at 100°C for 6h, then heat the oven to 160°C for 6h; 0.75g of sucrose, Dissolve 0.08g of concentrated sulfuric acid in 5.0g of water, immerse the above material into the mixture, dry it in an oven at 100°C for 6h, then heat the oven to 160°C for 6h; calcinate the obtained product at 900°C in a nitrogen atmosphere for 6h, The heating rate is 2.5°C / min, and finally the product is soaked in hydrofluoric acid, washed and dried. Finally, ordered mesoporous carbon material CMK-3 was obtained.

[0039] Immerse 300mg of CMK-3 nanomaterials into 15mL of 1.75mol / L ammonium persulfate solution and 15mL of 2mol / L sulfuric acid solution, stir in a water bath at 40°C for 24h, wash and dry to obtain carboxyl-functionalized mesoporous Carbon material O-CMK-3.

[0040] 5mg EDC and 6mg NHS were added to 30mL Tris-HCL buffer, 30mg carboxyl-func...

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Abstract

The invention discloses an electrochemical detection method of Staphylococcus aureus. The method comprises the following steps: preparing an ordered mesoporous carbon nano material CMK-3 by a template process, and carrying out surface carboxyl functionalization modification to obtain a carboxyl-functionalized mesoporous carbon nano material O-CMK-3; immobilizing sortase A (Srt A) onto the carboxyl-functionalized mesoporous carbon nano material O-CMK-3 by a chemical covalent binding technique to obtain an immobilized sortase nano composite material O-CMK-3-M; and preparing a carbon paste electrode from the immobilized sortase nano composite material, graphite, paraffin and other materials, immersing the carbon paste electrode into Staphylococcus aureus solutions with different concentrations, and detecting Staphylococcus aureus by using a tri-electrode detection system. Compared with the existing detection technique, the method has the advantages of short detection time and the like, is convenient to operate, is recyclable, and thus, has favorable application prospects.

Description

[0001] Technical field: [0002] The invention designs an electrochemical detection method for Staphylococcus aureus, belonging to the field of microbial detection. [0003] Background technique: [0004] Staphylococcus aureus, belonging to the genus Staphylococcus, is an important pathogen of humans, causing many serious infections. Typical Staphylococcus aureus is spherical, with a diameter of about 0.8um, arranged in clusters of grapes under the microscope. Staphylococcus aureus has no spores, flagella, most of them have no capsule, and Gram stain is positive. Its nutritional requirements are not high, and it grows well on ordinary medium. The optimum growth temperature is 37°C, the optimum growth pH is 7.4, and it has high salt tolerance. It is a kind of bacteria that is not conducive to human body. [0005] Staphylococcus aureus is ubiquitous in nature and can be found in air, water, dust, and human and animal waste. Therefore, there are many opportunities for food to b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327
CPCG01N27/3278
Inventor 王虹苏陈洋牛效迪吕卓席丽娟
Owner JILIN UNIV
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