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A Staphylococcus Phage and Its Application in Staphylococcus Infection

A staphylococcal infection, staphylococcus technology, applied in virus/phage, medical raw materials derived from virus/phage, application, etc., can solve problems such as difficulties in clinical treatment, improve economic benefits, improve skin, and reduce the emergence of drug-resistant strains The effect of the frequency

Active Publication Date: 2019-10-18
QINGDAO PHAGEPHARM BIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the past, antibiotics were more effective drugs for the treatment of this bacterial infection, but in recent years, the drug resistance of staphylococci has become stronger and stronger, especially the production of methicillin-resistant Staphylococcus aureus (MRSA), which has brought great impact to clinical treatment. Difficulties

Method used

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  • A Staphylococcus Phage and Its Application in Staphylococcus Infection
  • A Staphylococcus Phage and Its Application in Staphylococcus Infection
  • A Staphylococcus Phage and Its Application in Staphylococcus Infection

Examples

Experimental program
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Effect test

Embodiment 1

[0042] Screening and purification of embodiment 1 phage

[0043] 1. Recovery culture of staphylococcus

[0044] Pick the bacteria liquid of Staphylococcus, streak it on the ordinary agar plate, separate the single colony, and cultivate it in a constant temperature incubator at 37°C for 16-24h.

[0045] 2. Preparation of Staphylococcus Proliferation Solution

[0046] A single colony was picked, inoculated into a 5 mL LB broth test tube, and cultured overnight at 37° C. with shaking at 170 rpm to obtain a proliferation solution.

[0047] 3. Sample collection and processing

[0048] Take sewage and fur from pig farms, soak them in nutrient broth, incubate overnight at 37°C, take out 5mL of liquid, centrifuge at 10,000r / min for 10min, and filter the supernatant through a 0.22μm sterile microporous membrane to obtain phage stock solution , stored at 4°C.

[0049] 4. Isolation of phage

[0050] Isolate phages by double-layer plate method, dilute the stock solution of phages 10 ...

Embodiment 2

[0052] A single phage plaque was picked and placed in 1 mL of LB broth, and incubated in a 40°C water bath for 30 min to obtain a phage extract. Pick a single colony of Staphylococcus, inoculate in 5mL liquid LB medium, and cultivate overnight at 37°C with shaking at 170rpm; take 200μL phage extract and 200μL host bacteria proliferation solution in 5mL liquid LB medium, shake at 37°C and 170rpm until The liquid became clear, centrifuged at 10,000 rpm for 10 min, the supernatant was taken, and filtered through a 0.22 μm sterile microporous membrane to obtain a phage proliferation liquid. The biological characteristic of embodiment 2 bacteriophage

[0053] 1. Electron microscope observation of phage

[0054] Take the phage suspension purified by ultracentrifugation and drop a little on the copper grid covered with polyvinyl formaldehyde film, stain with 2% phosphotungstic acid for 5-10min, put the copper grid on dry filter paper, dry naturally, and use Hitachi HT7700 transmissi...

Embodiment 3

[0067] The safety test of embodiment 3 bacteriophage

[0068] Select 20 18-20g newly-adult Kunming mice (half male and half female) and randomly divide them into experimental group and blank control group (half male and female in each group). 9 PFU / mL and 0.2 mL of broth were fed continuously for 5 days, and the behavior and growth of the mice were observed.

[0069] It was found that the mice in the phage group and the control group grew in the same condition without adverse reactions, thus confirming that the phage SA1 was safe and had no toxic side effects.

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Abstract

The invention relates to staphylococcus bacteriophage and an application thereof in staphylococcal infection and particularly relates to staphylococcus bacteriophage. The staphylococcus bacteriophageis preserved in CGMCC, the preservation date of which is 27th, July, 2017, and the preservation number of CGMCC No. 14330. The bacteriophage has a relatively good splitting action on staphylococcus. The preparation can be used independently or used with cocktail to provide a safe, toxic and side effect and residual effect-free bacteriophage product source for treatment of staphylococcal infection.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a phage strain capable of lysing staphylococcus and the application of the phage in staphylococcus infection. Background technique [0002] Staphylococci are Gram-positive cocci. Staphylococcus is the most common pyogenic coccus, with a diameter of about 0.8 μm and a small spherical shape. According to different biochemical reactions and pigment production, Staphylococcus can be divided into Staphylococcus aureus, Staphylococcus epidermidis and Staphylococcus saprophyticus. Among them, Staphylococcus aureus is mostly pathogenic bacteria, Staphylococcus epidermidis is occasionally pathogenic, and Staphylococcus saprophyticus is generally not pathogenic. [0003] Staphylococcus can cause staphylococcal disease in chickens after infecting chickens. The clinical symptoms are acute septicemia symptoms, arthritis, skin necrosis and chick omphalitis. The disease occurs all the year round, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00A61K35/76A61P31/04A23K10/18C12R1/92
Inventor 任慧英孙虎芝刘爽潘强王倩闫艳新庄盈婷
Owner QINGDAO PHAGEPHARM BIO TECH CO LTD
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