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Biological reagent for treating carbapenems resistant acinetobacter baumannii infection

A technology of Acinetobacter baumannii and carbapenems, applied in biochemical equipment and methods, antibacterial drugs, plant genetic improvement, etc., can solve the problems of phage application limitations

Inactive Publication Date: 2013-01-09
重庆市人民医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This highly specific host selectivity greatly limits the application of phages

Method used

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  • Biological reagent for treating carbapenems resistant acinetobacter baumannii infection
  • Biological reagent for treating carbapenems resistant acinetobacter baumannii infection
  • Biological reagent for treating carbapenems resistant acinetobacter baumannii infection

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Embodiment 1

[0077] A. Isolation and purification of phage AB3;

[0078] Take sewage 1 from the hospital sewage treatment center, add 40 strains of carbapenem-resistant Acinetobacter baumannii suspension in the early logarithmic growth phase and 50ml of LB into the sewage at 37°C for 24h. Sterilize by filtration after centrifugation. After 10 μl of treated sewage was mixed with 200 μl of host bacteria for 20 minutes, it was mixed with 2 ml of top layer agar and plated. Uniform phage can be obtained after 5 repeated purifications of a single plaque, and the concentration of the obtained phage was confirmed by double-layer agar method. A total of 10 lytic carbapenem-resistant Acinetobacter baumannii phages were isolated. Make a lawn from 40 carbapenem-resistant Acinetobacter baumannii strains, take 10ul of each of the 10 isolated phages (108PFU / ml) and drop them on the corresponding positions on each lawn, and wait for drying at 37°C for 12 hours. The formation of phage plaques was observ...

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Abstract

The invention discloses a biological reagent for treating carbapenems-resistant acinetobacter baumannii infection. The reagent is a protein solution of lyase of a bacteriophage AB3. A preparation method includes separating and purifying the bacteriophage AB3, extracting a bacteriophage AB3 genome, subjecting deoxyribonucleic acid (DNA) of the bacteriophage AB3 to enzyme digestion analysis, performing sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of capsid protein of the bacteriophage AB3, amplifying a lyase gene of the bacteriophage AB3, constructing a prokaryotic expression carrier and expressing and purifying the lyase gene. By means of a method that the prokaryotic expression carrier is constructed to express and purify the lyase gene of the bacteriophage AB3, the biological reagent can be used for clinically treating the carbapenems-resistant acinetobacter baumannii infection, and the biological reagent has the advantages of being high in specificity, fast in effects and capable of being used as a novel antibacterial agent and has unique advantages and good application prospects.

Description

[0001] technical field [0002] The invention relates to the field of biology, in particular to a biological reagent for treating carbapenem-resistant Acinetobacter baumannii infection. Background technique [0003] The widespread resistance of Acinetobacter baumannii to commonly used antibacterial drugs makes the treatment of Acinetobacter baumannii increasingly difficult. Carbapenem antibiotics are considered to be one of the most effective drugs for the clinical treatment of Acinetobacter baumannii infection. In recent years, the proportion of carbapenem-resistant Acinetobacter baumannii infection has increased year by year. Once the bacteria are resistant to carbapenem antibiotics, they are basically resistant to other antibiotics, and patients often die because there is no drug available. Researchers turned their attention to anti-infection treatment to the study of phages. Phage is a kind of bacteria-dependent virus, also known as bacterial virus, which can rapidly pro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/51C12N15/60C12N15/70C12N9/88A61P31/04
Inventor 张劼李秀娟
Owner 重庆市人民医院
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