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Probe for detecting beta-lactamase and application of probe in fluorescence detection of drug-resistant bacteria

A technology of lactamase and drug-resistant bacteria, applied in the field of biochemical technology and medicine, can solve problems such as affecting the results of the test

Active Publication Date: 2020-09-08
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In complex environments with differences in temperature, humidity, etc. from the laboratory, the standard curve will no longer be applicable, which will affect the actual test results

Method used

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  • Probe for detecting beta-lactamase and application of probe in fluorescence detection of drug-resistant bacteria
  • Probe for detecting beta-lactamase and application of probe in fluorescence detection of drug-resistant bacteria
  • Probe for detecting beta-lactamase and application of probe in fluorescence detection of drug-resistant bacteria

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Embodiment 1, the preparation of fluorescent probe B1, P1, P2, P3

[0048] Synthetic route of probe B1

[0049]

[0050] Step 1: Synthesis of Compound B02

[0051] Cefalotin sodium (0.60±3.00g) was dissolved in water (4.00±2.00mL), and added to -20℃ 19% sodium hydroxide (6.00±3.00ml) aqueous solution at 0°C. The reaction mixture was stirred at -10°C for 135±5 seconds, then quenched by the rapid addition of glacial acetic acid (2.00±1.00 mL) at room temperature. Subsequently, the reaction vessel was transferred to an ice-water bath, and the pH of the mixture was immediately lowered to 1.5 by adding concentrated hydrochloric acid at -20°C. The filtered white solid was washed twice with water and freeze-dried to obtain compound B02 as a white powder. Wherein the name of compound B02 is: (6R, 7R)-3-(hydroxymethyl)-8-oxo-7-(2-(thiophen-2-yl)acetamide)-5-thia-1-azabicyclo [4.2.0] Oct-2-ene-2-carboxylic acid.

[0052] Step 2: Synthesis of compound B03

[0053] A solut...

Embodiment 2

[0112] Embodiment 2, the preparation of the fluorescent detection test paper of bacterial drug resistance and the detection experiment of bacterial drug resistance

[0113] Specific detection method

[0114] 1. Preparation of test strips

[0115] Step 1. Select a piece of chromatographic paper as the sampling layer, and use software to draw the first layer of graphics on the chromatographic paper, use wax printing technology to form detection micro-regions on the chromatographic paper, and print the printed chromatographic The paper is baked in an oven, so that the wax melts and penetrates the thickness of the paper to form a hydrophilic-hydrophobic barrier; the area connected by the two circles shown in the sample inlet area and the sample penetration 0 area is the hydrophilic area, and the rest Some are wax-printed hydrophobic areas.

[0116] Step 2. Select another piece of paper as the detection layer, and use software to draw the second layer of graphics on the chromatogra...

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Abstract

The invention discloses a probe for detecting beta-lactamase and application of the probe in fluorescence detection of drug-resistant bacteria. According to the detection method, whether drug-resistant bacteria exist or not (positive reference) is distinguished through a fluorescent probe B1; three different types of beta lactamases A, B and C are detected through fluorescent probes P1, P2 and P3.Reactants of the four probes are different, fluorescence reporter groups are the same, strong fluorescence signals are generated after beta-lactamase substrates are hydrolyzed, so that the bacterialdrug resistance in a sample is detected. The fluorescence detection method not only can distinguish drug-resistant / non-drug-resistant bacteria, but also can judge the drug resistance type and multipledrug resistance conditions of the bacteria. Detection test paper corresponding to the detection method provided by the invention has the characteristic of multi-environment adaptation, and still hasquantitative detection capability in complex environments such as temperature / humidity difference and the like. Therefore, guarantee is provided for monitoring clinical multi-drug resistance and a method is also provided for designing quantitative detection test paper in complex environments such as fields, battlefield medical treatment and the like.

Description

technical field [0001] The invention relates to the fields of biochemical technology and medical technology, in particular to a probe for detecting beta-lactamase and its application in fluorescent detection of drug-resistant bacteria. Background technique [0002] Antibiotics are one of the most successful types of drugs in modern medicine, and their emergence and development are also one of the important milestones in modern medicine. However, due to the increase of people's income around the world, the way to obtain antibiotics has become easier and simpler, which has greatly increased the use of antibiotics, which is one of the important reasons for bacterial drug resistance. At the same time, the misuse and abuse of antibiotics have also exacerbated bacterial resistance. The above and other reasons have caused more and more infections caused by drug-resistant bacteria, and the probability of incurable infections is increasing. WHO's first global report on antibiotic r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6428G01N2021/6432G01N2021/6439
Inventor 李林丁阳吴琼余昌敏姬文辉黄晓郑冰杨乃娣于海东刘金华黄维
Owner NANJING UNIV OF TECH