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Electrochemical aptamer sensor for detecting staphylococcus aureus as well as preparation method and application method of electrochemical aptamer sensor

An aptamer sensor, Staphylococcus technology, applied in the field of biosensors, can solve the problems of poor clinical practicability, unreliable sensor response, instability, etc., and achieve good reproducibility, good selectivity, and appropriate thickness

Pending Publication Date: 2022-02-25
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, conventional biosensors have generally proven to have poor clinical utility, mainly due to unstable or unreliable sensor responses due to biological contamination (e.g., protein adsorption, platelet adhesion, and clot formation).

Method used

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  • Electrochemical aptamer sensor for detecting staphylococcus aureus as well as preparation method and application method of electrochemical aptamer sensor
  • Electrochemical aptamer sensor for detecting staphylococcus aureus as well as preparation method and application method of electrochemical aptamer sensor
  • Electrochemical aptamer sensor for detecting staphylococcus aureus as well as preparation method and application method of electrochemical aptamer sensor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] An electrochemical aptamer sensor for detecting Staphylococcus aureus and its preparation method, such as figure 2 shown, including the following steps:

[0066] (1) Immerse a double-electrode system consisting of a glassy carbon electrode and a platinum wire in a chitosan solution, and deposit it by electrochemical deposition to obtain a glassy carbon electrode modified by a chitosan film.

[0067]Chitosan was dissolved using 1M HCl solution, and then its pH was adjusted to 5-6 with 1M NaOH solution. Stir overnight with a magnetic stirrer, and filter the chitosan solution to remove undissolved particles. Polish the glassy carbon electrode with alumina powder with a particle size of 1.0 μm, 0.3 μm, and 0.05 μm, for three minutes each time. Sonicate the glassy carbon electrode in distilled water for 10 minutes, and then wash it with aqua regia (H 2 SO 4 / 30%H 2 o 2 7:3, V / V) for 10 minutes, and finally rinsed with water and ethanol for 3 times. The glassy carbon ...

Embodiment 2

[0085] A detection application method of an electrochemical aptamer sensor for detecting Staphylococcus aureus in PBS solution. The electrochemical aptasensor prepared in Example 1 is used for detection, including the following steps:

[0086] 1) Establish a standard curve equation according to the CV curve drawn by the cyclic voltammetry of the Staphylococcus aureus serial concentration standard solution:

[0087] A: Fc-Ru in an electrochemical aptasensor 3+ Add a known concentration of Staphylococcus aureus PBS solution to the dual-electron medium solution (50 μM in 0.1M PBS, pH 7.4); use cyclic voltammetry to measure the known concentration of Staphylococcus aureus solution, and scan the CV The voltage is from -0.5V to +0.5V, record the CV curve until the signal is stable; record the peak value of the Fc oxidation current of the Staphylococcus aureus solution at this concentration when the signal is stable;

[0088] B: Set the concentration range to 1-10 9 Repeat step A ...

Embodiment 3

[0095] A detection application method of an electrochemical aptamer sensor for detecting Staphylococcus aureus in a whole blood sample. The electrochemical aptasensor prepared in Example 1 is used for detection, including the following steps:

[0096] 1) Establish a standard curve equation according to the CV curve drawn by the cyclic voltammetry of the Staphylococcus aureus serial concentration standard solution:

[0097] A: Fc-Ru in an electrochemical aptasensor 3+ Add a known concentration of whole blood sample Staphylococcus aureus solution to the dual-electron medium solution (each 50 μM in 0.1M PBS, pH7.4); Measurement, CV scanning voltage from -0.5V to +0.5V, record the CV curve until the signal is stable; record the Fc oxidation current peak value of the Staphylococcus aureus solution at this concentration when the signal is stable;

[0098] B: Set the concentration range to 1-10 8 Repeat step A for different concentrations of S. aureus solutions in cfu / mL, and deri...

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Abstract

The invention provides an electrochemical aptamer sensor for detecting staphylococcus aureus as well as a preparation method and an application method of the electrochemical aptamer sensor. The preparation method comprises the following steps: (1) depositing chitosan on a glassy carbon electrode to obtain a chitosan film modified glassy carbon electrode; (2) depositing catechol on the chitosan film modified glassy carbon electrode to obtain a catechol-chitosan film modified glassy carbon electrode; and (3) finally, combining the glassy carbon electrode with the staphylococcus aureus aptamer to obtain the aptamer-catechol-chitosan film modified glassy carbon electrode. The prepared sensor is used for measuring current through a CV method, and the content of staphylococcus aureus in a to-be-detected sample is obtained according to a standard curve equation of an Fc oxidation current peak value and the staphylococcus aureus concentration. The electrochemical aptamer sensor disclosed by the invention has relatively high specificity, affinity and reproducibility, and has good selectivity and quick response characteristics; the interference of sulfenyl groups during whole blood sample detection can be overcome, the detection precision is high, and direct detection of clinical samples can be realized.

Description

technical field [0001] The invention relates to an electrochemical aptamer 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 is a common opportunistic 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. While mild skin and mucosal infections are usually self-limited, severe systemic infections are often associated with high mortality (20%-50%), high recurrence rates (5-10%), and pers...

Claims

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Application Information

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IPC IPC(8): G01N27/30G01N27/327G01N27/48
CPCG01N27/308G01N27/3277G01N27/48Y02A50/30
Inventor 刘正春陈伟来庆腾张燕科龙孟秋梁波
Owner CENT SOUTH UNIV