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Acinetobacter baumannii bacteriophage SH-Ab15519 and application thereof

A technology of Acinetobacter baumannii and sh-ab15519, applied in the field of microbiology medicine, can solve the problems of high specificity requirements of host bacteria and the lack of promotion of bacteriophage, and achieve the effect of large clinical application potential.

Inactive Publication Date: 2017-05-24
SHANGHAI JIAOTONG UNIV SCHOOL OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the application of phages has not been promoted. In addition to the discovery and application of antibiotics mentioned above, another reason is that phages have too high requirements for the specificity of host bacteria.

Method used

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  • Acinetobacter baumannii bacteriophage SH-Ab15519 and application thereof
  • Acinetobacter baumannii bacteriophage SH-Ab15519 and application thereof
  • Acinetobacter baumannii bacteriophage SH-Ab15519 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Isolation and screening of phage SH-Ab15519

[0031] Centrifuge the sewage sample solution for 10 minutes; filter the supernatant through a filter membrane with a pore size of 0.22 μm, mix it with the bacterial solution cultivated to the logarithmic growth phase, and incubate overnight at 37°C; add a small amount of chloroform, centrifuge at 4°C for 10 minutes, and take the supernatant , and incubate with the strains cultivated to the logarithmic growth phase at 37°C for 10min, spread the plate by double-layer agar, and culture at 37°C overnight; the obtained phage plaques were purified three times for further research.

Embodiment 2

[0032] Example 2 Purification of phage SH-Ab15519

[0033]The phage and bacteria were mixed and cultured. On the next day, add 2% chloroform and let stand at 4°C for 1 hour, then centrifuge at 6500r / min for 15min at 4°C. Add DNase and RNase at a final concentration of 1ug / mL to the supernatant, and place at room temperature for 30min. Add NaCl to a final concentration of 0.5 M, and after 1 hour of ice-bath, centrifuge at 11,000 g for 10 minutes at 4°C. Take the supernatant, add solid PEG8000 to a final concentration of 10% (w / v), mix well and let stand at 4° C. overnight. Take the mixed solution from the previous day, centrifuge at 4°C, 8500r / min for 20min, discard the supernatant, and use SMBuffer (NaCl 5.8g / L, MgSO 4 ·7H 2 O 2g / L, gelatin 0.1g / L, Tris·HCl 50mL) resuspended. Add an equal amount of chloroform and mix lightly for 30 seconds, centrifuge at 4000g for 10 minutes, and take the supernatant. Purification of phage by cesium chloride density gradient method. Afte...

Embodiment 3

[0034] Example 3 Determination of phage SH-Ab15519 titer

[0035] The titer of phage——plaque forming unit (pfu)—was determined by double-layer agar plate method. The amplified phage liquid was serially diluted 10 times to 10 3 to 10 6 density range. Take 400 μL of phage dilutions and mix them in a test tube. After adsorption at room temperature for 10 min, observe the phages by the double-layer agar plate method and calculate the pfu, and repeat 3 times. Phage titer (pful / mL) = average number of plaques × dilution factor × 100.

[0036] The diameter of the plaque formed by SH-Ab 15519 infected host bacteria was about 7-8mm. The spots are round and transparent with halos outside. see figure 1 .

[0037] In addition, its in vitro lytic capacity see Figure 7 .

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Abstract

The invention discloses acinetobacter baumannii bacteriophage SH-Ab15519 and application thereof. Acinetobacter baumannii bacteriophage SH-Ab15519 is assigned the accession number CCTCC NO:M 2016653. The novel acinetobacter baumannii bacteriophage SH-Ab15519 is obtained by separation and purification and can peculiarly and effectively kill acinetobacter baumannii, have good in vivo and in vitro antibacterial effects on medicine-resistant acinetobacter baumannii, provide an experimental basis for clinically developing a preparation for preventing and treating infection by medicine-resistant acinetobacter baumannii and have great clinical application potential.

Description

technical field [0001] The present invention relates to the field of microbiology medicine, more specifically, relates to a novel Acinetobacter baumannii phage SH-Ab15519 and its application. Background technique [0002] Acinetobacter baumannii (Ab) is a common opportunistic pathogen in clinic. In recent years, the proportion of Ab infection in nosocomial infections in my country has increased significantly, and its resistance to antibacterial drugs has also increased year by year, and the proportion of multi-drug resistant bacteria has increased significantly. According to research, there are significant differences in the resistance rate of Ab to antimicrobials among different hospitals and different departments, and it mostly occurs in cardiac surgery, and lung infection is more common, which causes Bowman's pneumonia with a high mortality rate. In addition, Ab was highly resistant to common antimicrobials, such as cefoperazone / sulbactam, ampicillin / sulbactam, and piper...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A61K35/76A61P31/04
CPCC12N7/00A61K35/76C12N2795/00021
Inventor 何平胡付品罗婷婷徐梦莎
Owner SHANGHAI JIAOTONG UNIV SCHOOL OF MEDICINE
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