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Method for detecting drug resistance of bacteria based on AIE fluorescent probe

A fluorescent probe, fluorescence detection technology, applied in fluorescence/phosphorescence, measurement device, material analysis by optical means, etc., can solve the problems of limited applicable drugs and strains, determination of growth focus, complicated operation, etc. High performance and real-time performance, accurate bacterial drug detection, high luminous efficiency

Active Publication Date: 2019-12-31
东莞市厚街医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the traditional bacterial susceptibility test (AST) is a growth-based drug susceptibility test method, which depends on the growth of microorganisms. Therefore, it takes 24-48 hours or longer to reach the level that can be distinguished visually or by instruments.
However, it is difficult to accurately and reproducibly determine the growth focus of bacterial strains, coupled with the strong subjectivity of the result judgment, the inability to measure the MIC value (disc diffusion method), and limited applicable drugs and strains, it cannot bring timely clinical results. Feedback of results not only affects the precise medication of critically ill patients, but also greatly affects the clinical value of drug sensitivity testing
[0003] In order to avoid these problems, several other test methods have been reported, including fluorescein-labeled phage method, electrochemical detection method, real-time PCR method and method based on nanoparticles, etc., but these methods also have disadvantages: expensive instruments, High cost, complicated operation, low sensitivity and high repeatability limit the application of the above methods

Method used

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  • Method for detecting drug resistance of bacteria based on AIE fluorescent probe
  • Method for detecting drug resistance of bacteria based on AIE fluorescent probe
  • Method for detecting drug resistance of bacteria based on AIE fluorescent probe

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] See figure 1 , 4 , a method for detecting bacterial drug resistance based on AIE fluorescent probes, comprising the steps of:

[0048] Step (1): Prepare the Escherichia coli strain with physiological saline to a bacterial suspension with a McFarland turbidity of 0.5;

[0049] Step (2): Dilute 1mL of 6PD-DPAN fluorescent probe and 10μL of the bacterial suspension in step (1) into 10mL of MOPS buffer, shake well to prepare a mixture;

[0050] Step (3): Add 50 μL of the mixed solution prepared in step (2) to each well of the disposable sterile 96-well plate 11, and then add it to the first well of the disposable sterile 96-well plate 11 50 μL of MOPS buffer solution was prepared as a control sample, and 50 μL of antibacterial drug solutions of different concentrations were added to each well except the first well to prepare several samples to be tested, and the plate was sealed with a transparent film to detect and Record the initial fluorescence intensity value FRET of...

Embodiment 2

[0065] See figure 1 , 5 , a method for detecting bacterial drug resistance based on AIE fluorescent probes, comprising the steps of:

[0066] Step (1): The Klebsiella pneumoniae strain is prepared with physiological saline to a bacterial suspension with a McFarland turbidity of 0.5;

[0067] Step (2): Dilute 1mL of 6PD-DPAN fluorescent probe and 10μL of the bacterial suspension in step (1) into 10mL of MOPS buffer, shake well to prepare a mixture;

[0068] Step (3): Add 50 μL of the mixed solution prepared in step (2) to each well of the disposable sterile 96-well plate 11, and then add it to the first well of the disposable sterile 96-well plate 11 50 μL of MOPS buffer solution was prepared as a control sample, and 50 μL of antibacterial drug solutions of different concentrations were added to each well except the first well to prepare several samples to be tested, and the plate was sealed with a transparent film to detect and Record the initial fluorescence intensity valu...

Embodiment 3

[0082] See figure 1 , 6 , a method for detecting bacterial drug resistance based on AIE fluorescent probes, comprising the steps of:

[0083] Step (1): Prepare the Staphylococcus aureus strain with physiological saline to a bacterial suspension with a McFarland turbidity of 0.5;

[0084] Step (2): Dilute 1mL of 6PD-DPAN fluorescent probe and 10μL of the bacterial suspension in step (1) into 10mL of MOPS buffer, shake well to prepare a mixture;

[0085] Step (3): Add 50 μL of the mixed solution prepared in step (2) to each well of the disposable sterile 96-well plate 11, and then add it to the first well of the disposable sterile 96-well plate 11 50 μL of MOPS buffer solution was prepared as a control sample, and 50 μL of antibacterial drug solutions of different concentrations were added to each well except the first well to prepare several samples to be tested, and the plate was sealed with a transparent film to detect and Record the initial fluorescence intensity value FR...

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Abstract

The invention belongs to the technical field of micro-detection and relates to a method for detecting the drug resistance of bacteria based on an AIE fluorescent probe. The method comprises the following steps that: a to-be-detected bacterial strain is prepared into a bacterium suspension with McFarland turbidity of 0.5 by using normal saline; a 6PD-DPAN fluorescent probe and the bacterium suspension are sucked and diluted into an MOPS buffer solution, so that a mixed solution can be prepared; the mixed solution is added into each hole in a sterile 96-well plate, antibacterial drug solutions with different concentrations are added into all the holes except the first hole, so that a plurality of to-be-detected samples can be prepared, and initial fluorescence intensity values are detected and recorded; and the sterile 96-well plate with the to-be-detected samples is incubated, the fluorescence intensity values of the to-be-detected samples are detected and recorded each time the sterile96-well plate has been incubated for a period of time. The detection method disclosed by the invention is simple and easy to implement; and the aggregation-induced emission characteristic and high light stability of the 6PD-DPAN fluorescent probe are utilized, so that the dynamic process of the combination of the 6PD-DPAN fluorescent probe and the bacterial strain can be monitored in real time.

Description

technical field [0001] The invention relates to the technical field of trace detection, in particular to a method for detecting bacterial drug resistance based on an AIE fluorescent probe. Background technique [0002] Bacterial susceptibility testing (AST) is an important means to provide information on the susceptibility of clinical pathogens to different antibacterial drugs, and it is of great value in guiding clinical medication, monitoring treatment effects, discovering drug-resistant strains, and developing and screening new antibacterial drugs. However, the traditional antibacterial susceptibility test (AST) is a growth-based antimicrobial susceptibility test method, which depends on the growth of microorganisms. Therefore, it takes 24-48 hours or longer to reach the level that can be distinguished visually or by instruments. However, it is difficult to accurately and reproducibly determine the growth focus of bacterial strains, coupled with the strong subjectivity of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6428
Inventor 付文金邓任堂赖丽莎谢岭平张露
Owner 东莞市厚街医院
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