Method for separating and screening phage for degrading Pseudomonas aeruginosa biofilm

A Pseudomonas aeruginosa, biofilm technology, applied in biochemical equipment and methods, viruses/phages, microorganisms, etc., can solve problems such as difficulties in clinical treatment, inability to effectively treat and control infections with antibiotics, etc., to overcome drug resistance the effect of

Inactive Publication Date: 2011-09-28
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] For a long time, antibiotics have been widely used in the treatment of bacterial diseases, leading to the continuous emergence of drug-resistant strains, which has brought great difficulties to clinical treatment
Clinically, it is easy to isolate Pseudomonas aeruginosa strains with multidrug resistance or pan-drug resistance, resulting in the ineffective treatment and control of infections caused by Pseudomonas aeruginosa with the currently used antibiotics

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] The name of the degrading Pseudomonas aeruginosa biofilm phage obtained in this example is: C12, the deposit number: CGMCCNo.4249, and the depository unit: General Microbiology Center of China Committee for the Collection of Microorganisms, address: Beichen West Road 1, Chaoyang District, Beijing Courtyard No. 3, date of preservation: October 26, 2010. The host bacteria were also preserved at the same time as the phage was preserved.

[0038] (1) Collect samples from the soil in Taifeng Park, Taida District, Tianjin;

[0039](2) Amplify phage: 37 ℃, under the condition of 120rpm, culture host bacterium in test tube overnight, expand culture to logarithmic phase (OD600 about 0.5) with 1% inoculation amount in 250mL Erlenmeyer flask, add 5g sludge sample (or 5mL Sewage), after 48 hours of mixed culture, centrifuge at 8000g for 10 minutes to remove most of the bacteria and sample impurities, collect the supernatant, and filter it with a 0.22 μm filter membrane to steriliz...

Embodiment 2

[0050] Collect muddy water samples at the seaside of Bohai Bay; the rest of the steps are the same as in Example 1.

Embodiment 3

[0052] Sludge samples from swine biogas digesters were collected; the rest of the steps were the same as in Example 1.

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PUM

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Abstract

The invention relates to a phage for degrading a Pseudomonas aeruginosa biofilm. The phage is characterized in that: the phage is named C12, the collection number is CGMCC No. 4249, the collection unit is China General Microbiological Culture Collection Center located at No. 3, No. 1 Yard, Beichen West Road, Chaoyang District, Beijing City, and the collection date is October 26, 2010. The invention provides a method for amplifying, separating and preserving the phage for degrading Pseudomonas aeruginosa. The capacity of degrading the Pseudomonas aeruginosa biofilm by the phage is also determined, finally the pure phage C12 with high titer and high biofilm degradation capacity is obtained, and the degradation amount of the pure phage C12 reaches 85.3 percent; the phage can degrade biofilms and crack viable bacteria in the biofilms; the clinical application of the phage can be realized; treatment problems of medicine resistance and biofilms of the Pseudomonas aeruginosa are solved; and the method is a feasible biological method.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for separating and screening phages, in particular to a method for separating and screening phages that degrade Pseudomonas aeruginosa biofilms. Background technique [0002] Pseudomonas aeruginosa is an opportunistic pathogen that widely exists in various environments and is one of the important Gram-negative bacteria that cause nosocomial infections. The bacterium has strong adaptability and can form biofilms on various biotic and abiotic surfaces, including medical devices or human tissues, which are difficult to remove. Studies have found that 80% of Pseudomonas aeruginosa infections are caused by biofilms. Common infections include burn patients, urinary tract infections, catheter infections, otitis media, and corneal infections. [0003] For a long time, antibiotics have been widely used in the treatment of bacterial diseases, leading to the continuous emergence of drug...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00
Inventor 杨洪江李玲艳梁莉张晓梅朱会霞
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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