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Electrochemical luminescence biosensor for detecting drug resistance of bacteria and preparation method of biosensor

A bioluminescent, drug-resistant technology, applied in the field of electrochemiluminescence, can solve the problems of antibiotic abuse, high noise, increased bacterial resistance, etc., and achieve the effects of high sensitivity, high sensitivity, and less reagents

Active Publication Date: 2020-07-28
NORTHWEST UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The direct detection method uses spectrophotometry to directly detect the hydrolysis of β-lactam in cell extracts, which is time-consuming and laborious and therefore cannot be used for routine detection
Matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) has been widely used in the study of antimicrobial resistance mechanisms in recent years, but relatively small antibiotics (~1000Da) are difficult to analyze because of their interaction with the matrix. Interactions create higher noise levels
In addition, the price of the instrument and its maintenance costs are also additional issues
Therefore, in the face of unclear whether the patient is infected with drug-resistant bacteria, doctors often use empiric therapy, which leads to the abuse of antibiotics and the increase of bacterial resistance

Method used

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  • Electrochemical luminescence biosensor for detecting drug resistance of bacteria and preparation method of biosensor
  • Electrochemical luminescence biosensor for detecting drug resistance of bacteria and preparation method of biosensor
  • Electrochemical luminescence biosensor for detecting drug resistance of bacteria and preparation method of biosensor

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preparation example Construction

[0040] A preparation method of electrochemiluminescence biosensor for detecting bacterial drug resistance, taking the detection of whether bacteria express MβLs as an example, the selected bacteria is E.coli BL21 / NDM-1E.coli. The specific preparation steps are as follows:

[0041] Step 1. Preparation of NH 2 -MIL-53(Al) nanosheets;

[0042] AlCl 3 ·6H 2 O (3mmol, 0.7243g) was dissolved in 15mL of deionized water, and 2-amino-1,4-phthalic acid (3mmol, 0.5435g) was slowly added under stirring conditions, and stirring was continued for another 30 minutes. Subsequently, 15mL of urea (6mmol, 0.3604g) aqueous solution was added dropwise to the above mixed solution and stirring continued for 30 minutes. After that, the above-mentioned mixture was transferred to a 50mL polytetrafluoroethylene autoclave, and the reaction was allowed to stand at 150℃ for 5h. After the completion of the reaction, the mixture was slowly cooled to room temperature, suction filtered to obtain a milky white and ...

Embodiment 1

[0068] A preparation method of electrochemiluminescence biosensor for detecting bacterial resistance, taking the detection of whether bacteria express MβLs gene as an example, the selected bacteria is E.coli BL21 / NDM-1E.coli, and the specific preparation steps are as follows:

[0069] Step 1. Preparation of NH 2 -MIL-53(Al) nanosheets;

[0070] AlCl 3 ·6H 2 O (3mmol, 0.7243g) was dissolved in 15mL deionized water, and slowly added 2-amino-1,4-phthalic acid (NH 2 -H 2 BDC, 3mmol, 0.5435g), and then continue to stir for 30 minutes. Subsequently, 15mL of urea (6mmol, 0.3604g) aqueous solution was added dropwise to the above mixed solution and stirring continued for 30 minutes. After that, the above-mentioned mixture was transferred to a 50mL polytetrafluoroethylene autoclave, and the reaction was allowed to stand at 150℃ for 5h. After the completion of the reaction, the mixture was slowly cooled to room temperature, suction filtered to obtain a milky white and slightly yellow precipi...

Embodiment 2

[0078] A preparation method of electrochemiluminescence biosensor for detecting bacterial drug resistance, taking the detection of whether bacteria express MβLs gene as an example, the selected bacteria is not E.coli BL21 / NDM-1E.coli, the specific preparation steps are as follows:

[0079] Step 1. Preparation of NH 2 -MIL-53(Al) nanosheets;

[0080] AlCl 3 ·6H 2 O (3mmol, 0.7243g) was dissolved in 15mL deionized water, and slowly added 2-amino-1,4-phthalic acid (NH 2 -H 2 BDC, 3mmol, 0.5435g), and then continue to stir for 30 minutes. Subsequently, 15mL of urea (6mmol, 0.3604g) aqueous solution was added dropwise to the above mixed solution and stirring continued for 30 minutes. After that, the above-mentioned mixture was transferred to a 50mL polytetrafluoroethylene autoclave, and the reaction was allowed to stand at 150℃ for 5h. After the completion of the reaction, the mixture was slowly cooled to room temperature, suction filtered to obtain a milky white and slightly yellow pr...

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Abstract

The invention discloses an electrochemical luminescence biosensor for detecting drug resistance of bacteria and a preparation method of the electrochemical luminescence biosensor. The preparation method comprises the following steps of: preparing an NH2-MIL-53 (Al) nanosheet; preparing a probe solution for detecting the drug resistance of the bacteria; and assembling the electrochemical luminescence biosensor for detecting the drug resistance of the bacteria. The electrochemical luminescence biosensor provided by the invention can rapidly, simply, reliably and effectively detect drug resistance. An electrochemical luminescence technology is used as a signal output mode; the technology has extremely high sensitivity; and the electrochemical luminescence biosensor detects whether the bacteria have drug resistance or not and can be used for quantitatively detecting the concentration of escherichia coli.

Description

Technical field [0001] The invention relates to the field of electrochemiluminescence technology, in particular to an electrochemiluminescence biosensor for detecting bacterial drug resistance and a preparation method thereof. Background technique [0002] The resistance of pathogenic bacteria to antibiotics continues to increase. According to media reports, "super bacteria" have gradually attracted public attention. New Delhi metal-β-lactamase-1 (NDM-1) is a drug-resistant gene carried by super bacteria. It can neutralize β-lactam antibiotics and make bacteria resistant to almost all antibiotics. The extremely strong drug resistance of NDM-1 super bacteria greatly increases the difficulty in the treatment of infections caused by it. Common drug-resistant bacteria cannot "pass" drug-resistant genes to other bacteria, while NDM-1 genes can be different Transfer between bacteria to transmit resistance. The currently discovered NDM-1 mainly exists in Escherichia coli and Klebsiell...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/569G01N33/531G01N21/76G01N27/26
CPCG01N21/76G01N27/26G01N33/531G01N33/56911G01N33/68G01N2333/4724G01N2400/50
Inventor 马芬刘家玮陈玉孙利娜
Owner NORTHWEST UNIV
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