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A kind of electrochemical detection method of bacterial drug resistance

An electrochemical and drug resistance technology, applied in the direction of biochemical equipment and methods, electrochemical variables of materials, measurement/inspection of microorganisms, etc., can solve the problems of complex instruments, long detection time, mixed steps, etc. Simple, low-cost, fast results

Active Publication Date: 2021-01-29
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In order to realize rapid and accurate detection of bacterial drug resistance, the present invention constructs a method based on electrochemical detection, which can quickly, accurately and non-specifically detect bacterial drug resistance, and solve the problem of long detection time and problems in the prior art of drug resistance detection. Mixed steps and complicated technical problems of required instruments

Method used

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  • A kind of electrochemical detection method of bacterial drug resistance
  • A kind of electrochemical detection method of bacterial drug resistance
  • A kind of electrochemical detection method of bacterial drug resistance

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preparation example Construction

[0031] Preparation of modified screen-printed electrodes:

[0032] (1) First modify the multi-walled carbon nanotubes on the surface of the screen-printed electrode. The modification method: activate the screen-printed electrode in 0.5mol / L iron-sulfuric acid solution, scan by cyclic voltammetry, and the voltage is -0.2V ~0.6V, scan rate: 50mv / s, scan cycle: 10; configure 1mg / mL multi-walled carbon nanotube solution, ultrasonic for 30min, after it is mixed evenly, drop-coat it on the surface of the activated screen printing electrode , let it dry naturally at room temperature;

[0033](2) Modify the nano-gold particles by electroplating, the modification method: immerse the multi-walled carbon nanotube screen-printed electrode in 10mL of gold-plating solution containing 1% chloroauric acid and 1mmol / L sulfuric acid solution for gold plating, using time current method, electroplating voltage: -0.3V, electroplating time: 100s; finally, immerse the screen-printed electrode in aq...

Embodiment 1

[0035] (1) Bacteria treatment:

[0036] Salmonella typhimurium ATCC14028 was cultured with NB medium in a shaker at 37°C, and then the turbidity was measured with a bacterial turbidity meter after deducting the background, and then converted into the original concentration of the bacterial solution. According to the original concentration, the bacterial solution was diluted to 7.5×10 with NB medium 5 cfu / mL. Then add resazurin and different amounts of antibiotic ofloxacin to mix, shake and incubate in a metal bath at 37°C for 4 hours; the concentration of resazurin in the solution is 0.2mmol / L, and the concentration of antibiotic ofloxacin is respectively 128, 64, 32, 16, 8, 4, 2, 1, 0.5, 0.25, 0.125 μg / mL. At the same time, prepare an identical bacterial solution for control experiments without antibiotics.

[0037] (2) Drug resistance detection:

[0038] To detect bacterial resistance, the above-incubated solution was filtered through a 0.22 μm filter to remove bacteria,...

Embodiment 2

[0041] (1) Bacterial treatment:

[0042] Staphylococcus aureus ATCC6538 was cultured with NB medium in a shaker at 37°C, and then the turbidity was measured with a bacterial turbidity meter after deducting the background, and then converted into the original concentration of the bacterial solution. According to the original concentration, the bacterial solution was diluted to 7.5×10 with NB medium 5 cfu / mL. Then add resazurin and different amounts of antibiotic ofloxacin to mix, shake and incubate in a metal bath for 4 hours at 37°C; the concentration of resazurin in the solution is 0.2mmol / L, and the concentration of antibiotic ofloxacin is 128, 64, 32, 16, 8, 4, 2, 1, 0.5, 0.25, 0.125 μg / mL. At the same time, prepare an identical bacterial solution for control experiments without antibiotics.

[0043] (2) Drug resistance detection:

[0044] To detect bacterial resistance, the above-incubated solution was filtered through a 0.22 μm filter to remove bacteria, and then the ...

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Abstract

The invention discloses an electrochemical detection method for bacterial drug resistance, which belongs to the technical field of electrochemical chips and bacterial detection. The invention utilizes a specific resazurin-containing sample solution in a specific electrochemical environment and combines differential pulse voltammetry to detect the drug susceptibility of bacteria. The method of the present invention can directly detect bacterial activity, has high accuracy, is simple to operate, can carry out the determination of drug resistance and minimum inhibitory concentration in a short time, and can be widely used in various researches and clinical detections in the fields of medicine and the like. .

Description

technical field [0001] The invention belongs to the technical field of electrochemical chips and bacteria detection, and in particular relates to an electrochemical detection method for bacterial drug resistance. Background technique [0002] Resazurin (RZ), also known as sodium sky blue, resin azure, is a redox dye, dark blue or black, with green luster, and is a safe and non-toxic water-soluble dye. As a redox indicator, in cell biology research, it is commonly used in the functional detection of mitochondria (resazurin method) and the assessment of cell viability and cell proliferation ability or for the detection and analysis of bacteria; as an anaerobic indicator, it can be used in Preparation of microbial culture medium. At the same time, as a redox mediator, resazurin can regulate the metabolic activities of microorganisms. The active substance with metabolic capacity irreversibly reduces resazurin (blue) to resorufin (pink), which continues to be reversibly reduced...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/48C12Q1/04
CPCC12Q1/04G01N27/48Y02A50/30
Inventor 孙秀兰任奕婧纪剑孙嘉笛皮付伟张银志
Owner JIANGNAN UNIV
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