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Microelectrode sensor for detecting staphylococcus aureus as well as preparation method and application method of microelectrode sensor

A staphylococcus and microelectrode technology, applied in the field of biosensors, can solve problems such as being unsuitable for bedside detection, expensive instruments, and high professional requirements, and achieve the effects of increasing electron exchange capacity, wide linear range, and high specificity

Active Publication Date: 2022-03-01
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The gold standard for the detection of S. aureus is still the culture method, but it is very time consuming, usually taking 1-2 days to obtain a single colony, and then 1-2 days to obtain the chemical identification and susceptibility results
In recent years, some rapid and automated detection methods have been developed, such as enzyme-linked immunosorbent assay (ELISA), polymerase chain reaction (PCR), flow cytometry, and mass spectrometry, although results can be obtained in only 1-5 hours, However, these methods require expensive instruments, complicated sample preparation, and high professional requirements, and are not suitable for remote and poor areas, nor are they suitable for point-of-care testing

Method used

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  • Microelectrode sensor for detecting staphylococcus aureus as well as preparation method and application method of microelectrode sensor
  • Microelectrode sensor for detecting staphylococcus aureus as well as preparation method and application method of microelectrode sensor
  • Microelectrode sensor for detecting staphylococcus aureus as well as preparation method and application method of microelectrode sensor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] A microelectrode sensor for detecting Staphylococcus aureus, the microelectrode sensor is a three-electrode system sensor, the counter electrode is a platinum wire electrode, the reference electrode is an Ag / AgCl electrode, and the working electrode is chitosan, phthalate Silver microelectrodes modified with phenol and Staphylococcus aureus aptamers.

[0043] In this embodiment, the working electrode is based on a silver microelectrode, chitosan is coated on the surface of the silver microelectrode, and catechol and Staphylococcus aureus aptamer are respectively grafted onto the chitosan.

[0044] In this example, based on the principle of microcapacitance, a modified layer was prepared on the surface of the electrode to amplify the detection signal; an ultrasensitive microelectrode sensor for the detection of Staphylococcus aureus was constructed. The electrode modification process is the deposition of chitosan, grafting of catechol and immobilization of aptamers. Sil...

Embodiment 2

[0049] A kind of preparation method of the microelectrode sensor that detects staphylococcus aureus in preparation embodiment 1, such as image 3 shown, including the following steps:

[0050] (1) A silver microelectrode is used as the working electrode, and a platinum wire is used as the auxiliary electrode to construct a dual-electrode system. Chitosan is electrochemically deposited on the surface of the working electrode to obtain a Chi-Ag electrode;

[0051] (2) the Chi-Ag electrode obtained in step (1) is used as a working electrode, with Ag / AgCl as a reference electrode, and platinum wire as a counter electrode, electrochemically grafting catechol on the surface of the Chi-Ag electrode; By applying a voltage of 0.6V for 500s through a CHI660e electrochemical workstation, catechol was grafted onto chitosan; a Cat-Chi-Ag electrode was obtained. The Cat-Chi-Ag electrode was washed with ultrapure water and dried with nitrogen flow;

[0052] (3) Soak the Cat-Chi-Ag electrod...

Embodiment 3

[0056] A detection method based on the microelectrode sensor detecting Staphylococcus aureus in embodiment 1, comprises the following steps:

[0057] 1) Fc-Ru in microelectrode sensor 3+ / 2+ Add the concentration standard solution of Staphylococcus aureus series into the dual-electron medium solution, use cyclic voltammetry at room temperature to obtain the peak current of the corresponding silver electrode specific response signal of each concentration standard solution, according to the concentration of Staphylococcus aureus and The relationship between the peak currents establishes the standard curve equation;

[0058] 2) Take or prepare the sample solution to be tested;

[0059] 3) adopting cyclic voltammetry to detect the peak current of the characteristic peak to be measured for the sample solution to be tested in step 2);

[0060] 4) Bring the peak current obtained in step 3) into the standard curve equation in step 1) to calculate the concentration of Staphylococcus a...

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Abstract

The invention provides a microelectrode sensor for detecting staphylococcus aureus as well as a preparation method and an application method of the microelectrode sensor. The microelectrode sensor is a three-electrode system sensor, a counter electrode of the microelectrode sensor is a platinum wire electrode, a reference electrode of the microelectrode sensor is an Ag / AgCl electrode, and a working electrode of the microelectrode sensor is a silver microelectrode modified by chitosan, catechol and a staphylococcus aureus aptamer. The microelectrode sensor disclosed by the invention is constructed on the basis of a microelectrode with a micro-nano structure on the surface, double media (Fc-Ru < 3 + >), chitosan and catechol. The working electrode of the microelectrode sensor can specifically capture staphylococcus aureus. In the CV test process of the microelectrode sensor, CI <-> in bacteria can leak to the surface of the working electrode under the action of local high field intensity, and when CI <-> ions exist, the cathode reduction peak of the working electrode between 0 and-0.1 V can be specifically enhanced, so that a linear relation between bacterial concentration increase and current signal enhancement is established; the purpose of specifically detecting bacteria is achieved.

Description

technical field [0001] The invention relates to a microelectrode sensor for detecting Staphylococcus aureus, a preparation method and an application method thereof, and belongs to the technical field of biosensors. Background technique [0002] Staphylococcus aureus was first discovered and named by Dr. Alexander Ogston in 1880. Typical Staphylococcus aureus is spherical, grape-like clusters, about 0.8 μm in diameter, and Gram staining is positive. Staphylococcus aureus is a common pathogen that commonly colonizes human skin and mucosal surfaces, especially the anterior nares (approximately 30% of the general population). When the host's immunity is weakened or the skin and mucosal barriers are disrupted, it can enter any organ or enter the blood, causing skin and soft tissue infections (impetigo, folliculitis, and scalded skin syndrome) in mild cases and severe systemic diseases such as bacteremia, endocarditis, osteomyelitis, hemolytic pneumonia, and toxic shock syndrome...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327G01N27/48
CPCG01N27/3278G01N27/48G01N27/3277G01N27/3275
Inventor 刘正春来庆腾牛奇斌陈伟张燕科王福亮龙孟秋梁波
Owner CENT SOUTH UNIV
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