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Pseudomonas aeruginosa phage and application thereof

A Pseudomonas aeruginosa, phage technology, applied in phage, virus/phage, application and other directions, can solve problems such as untimely treatment and ineffective medication

Active Publication Date: 2019-08-20
QINGDAO PHAGEPHARM BIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The present invention provides a bacteriophage that can be injected into the blood through oral administration, intramuscular injection, and intraperitoneal injection and its application, aiming to solve the problem of untimely treatment of mink hemorrhagic pneumonia due to the acute onset of mink hemorrhagic pneumonia, as well as the drug resistance of pathogenic bacteria. Difficulties such as ineffective medication

Method used

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  • Pseudomonas aeruginosa phage and application thereof
  • Pseudomonas aeruginosa phage and application thereof
  • Pseudomonas aeruginosa phage and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] The isolation culture and biological characteristic of embodiment 1 bacteriophage

[0032] (1) Recovery culture of strains and preparation of bacterial suspension

[0033] Pick the frozen bacterial solution of Pseudomonas aeruginosa and streak it in three areas on the NAC plate, separate single colonies, and culture them in a 37°C incubator for 16-24 hours. Pick a single colony, inoculate into 5ml NB broth, and culture in an air shaker at 37°C at 170rpm for 16h to obtain a single bacterial suspension.

[0034] (2) Isolation and purification of phage

[0035] Take feces samples, litter, sewage, fur and other samples and put them into conical flasks filled with NB broth. In an air shaker, shake culture at 170rpm overnight, centrifuge at 10000rpm for 5min, and filter to sterilize with a 0.22μm bacterial filter. Mix the filtrate and the host bacteria, incubate at 37°C for 5 minutes, pour the double-layer plate, put it into a 37°C incubator and incubate it upside down aft...

Embodiment 2

[0054] The mensuration of embodiment 2 phage lysis spectrum and in vitro lysis test

[0055] (1) Determination of phage lysis profile

[0056] The single-spot method was used to determine the lysis profile of the phage, and the steps were as follows: the bacterial suspension of the host bacteria was obtained according to the method in Example 1, and 51 strains of Pseudomonas aeruginosa were respectively derived from mink, fox, cattle, pigs, poultry, rabbits, etc. animal. Add 100 μl of bacterial suspension to the upper layer of agar to prepare a double-layer plate. After the agar solidifies, take 1 μl of phage proliferation solution and drop it on the plate. After natural drying, place it in an incubator at 37°C and incubate it upside down for 6-8 hours. Observation results.

[0057] The results showed that 51 strains of Pseudomonas aeruginosa were used as host bacteria to measure the lysis spectrum, and phage ASP11 could lyse 44 of them, with a lysis rate of 86.27%.

[0058...

Embodiment 3

[0066] The safety test of embodiment 3 bacteriophage

[0067] Select 12 healthy BALB / C mice with a body weight of 18-20 g, 20 males and females, and divide them into experimental group and control group. The mice in each group are divided into male and female mice, respectively, and fed with purified phage proliferation solution (200 μl 10 10 PFU / ml) and normal saline (200 μ l), fed continuously for 7 days, observed the behavioral performance of the mice, and observed the changes of the organs of the mice by autopsy.

[0068] The results showed that the mice in the experimental group and the control group had no abnormal behavior, and the liver, lungs, heart, spleen, kidneys and other organs were normal after autopsy, and there was no significant difference from the control group.

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Abstract

The present invention relates to a pseudomonas aeruginosa phage and an application thereof, and particularly discloses the phage capable of entering blood and the application thereof. The phage singlebody is the pseudomonas aeruginosa phage with a Latin name P. Aeruginosaphage, and named as ASP11, and has a broad-spectrum bactericidal ability against pseudomonas aeruginosa. The phage ASP11 is preserved in China General Microbiological Culture Collection Center, a preservation date is September 26, 2018 and a preservation number is CGMCC NO.16395. Both in vivo and in vitro tests show that thephage has a strong cleavage effect on the pseudomonas aeruginosa and provides an effective method for prevention and treatment of pseudomonas aeruginosa infected diseases.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a phage isolate capable of specifically lysing Pseudomonas aeruginosa strains and an application thereof. Background technique [0002] Mink hemorrhagic pneumonia is an acute infectious disease caused by Pseudomonas aeruginosa. It has a rapid onset and is difficult to control. It was endemic in many countries and regions, resulting in the death of a large number of minks and serious economic losses. Pseudomonas aeruginosa has a variety of natural and acquired mechanisms to resist a variety of antibiotics. These mechanisms include efflux pumps, antibiotic degradation, modification of enzyme systems, and the function of reducing membrane permeability. Many strains of Pseudomonas aeruginosa that are drug resistant and clinically isolated show simultaneous resistance to multiple antibiotics. Meat feed contaminated with Pseudomonas aeruginosa, feces, urine, secretions of infe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A23K10/18A23K50/40A23L33/10A61K35/76A61P31/04
CPCA23K10/18A23K50/40A23L33/10A23V2002/00A61K35/76A61P31/04C12N7/00C12N2795/00021A23V2200/30
Inventor 潘强任慧英孙虎芝刘爽闫艳新
Owner QINGDAO PHAGEPHARM BIO TECH CO LTD
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