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Light green balloon bacteriophage AVP and use thereof

A light green, phage technology, applied in the field of microorganisms, to achieve the effect of strong killing activity and wide cracking spectrum

Active Publication Date: 2019-01-25
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, most phage biological control studies focus on the control of Escherichia coli, Listeria, Staphylococcus aureus, and Clostridium, and phages that can specifically control A. viridans have not been reported yet.

Method used

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  • Light green balloon bacteriophage AVP and use thereof
  • Light green balloon bacteriophage AVP and use thereof
  • Light green balloon bacteriophage AVP and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] The phage of the present invention is a novel phage isolated from sewage, and the phage has an icosahedral head and a retractable tail; the phage can form translucent plaques on 1.5% BHI agar medium without halos around the edges, Clear and regular, with a diameter of 0.1-1mm; the restriction map of genomic nucleic acid shows that the phage nucleic acid is double-stranded DNA (dsDNA); it has been preserved in the China Center for Type Culture Collection on March 25, 2018, and the preservation number is CCTCC NO: M 2018155, Address: Wuhan University, Wuhan, China.

Embodiment 2

[0028] Phage isolation and preparation

[0029] The isolation procedure of phage is described in detail below. The fecal liquid sewage sample in the present invention is collected from Huoshao Li Sewage Ditch in Changchun City; Collect sewage, filter it with gauze, or centrifuge at 6000r / min for 10min, and take the supernatant; use the treated sewage instead of ddH2O to prepare BHI medium (100mL); add 1mL of overnight cultured bacteria to the medium, and incubate at 37°C for 10-12h; Take 1 mL of the culture, centrifuge at 12,000 r / min for 5 min, filter the supernatant with a 0.22 μm filter and save it, and use the obtained filtrate for a spot test to check whether it includes phages capable of killing Coccus aureus.

[0030] Plaque test was carried out as follows: inoculate Coccus aureus AV-XHY in BHI liquid medium at a ratio of 2%, and cultivate overnight at 37°C with shaking. Take 100 μL (OD 600 =1.5) of the bacterial culture solution prepared above and spread it on the BH...

Embodiment 3

[0033] Phage amplification and purification

[0034]On the double-layer plate forming phage plaques, use the tip of a sterile pipette to pick up a single phage plaque with a large diameter, relatively round and translucent, inoculate it in 5 ml BHI liquid medium, and add phage 200 μL of host bacterial solution, mix well, act at room temperature for 15 minutes, incubate at 37°C for 10-14 hours, centrifuge at 12,000 rpm, 4°C for 10 minutes, and take the supernatant; repeat the double-layer plate experiment, and pick a single phage plaque repeatedly in this way for 4-5 times. Phage were purified into plaques of equal size.

[0035] Take 1 mL of freshly cultured host bacteria and add 300 μL of phage lysate (the ratio of a single phage culture to host bacteria is 1:1, 1:10 and 1:100, respectively). Incubate at 37°C for 20 minutes to make the phage particles adsorb to the host bacteria; add 800mL BHI liquid medium, then add CaCl 2 The mother solution was grown to a final concentr...

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Abstract

A light green balloon bacteriophage AVP is brought forward, deposited in China typical culture preservation center with that deposit number of CCTCC NO:M 2018155, deposited on 25 March 2018. The new light green balloon bacteriophage AVP can lyse the bacteriophage AVP lysate of Aeromonas aeruginosa, the bacteriophage has a wide lysis spectrum, not only can it lyse the pale green balloon bacteria, It can also lyse Staphylococcus aureus and Staphylococcus epidermidis. It can lyse standard strains or clinical strains isolated from environment or diseased animal feces. It can be used alone or in combination with other substances to provide a safe and non-toxic bacteriophage sterilization product for disinfection and purification of environment.

Description

technical field [0001] The present invention relates to a bacteriophage isolated from nature that infects and kills A. viridans, and further discloses a new phage AVP of A. viridans, and also provides its use, and uses the phage lysate as an active The invention relates to a method for cracking coccus aureus and staphylococcus with a combination of ingredients, belonging to the technical field of microorganisms. Background technique [0002] Light green balloon ( Aeroeoccus viridans ) in the family Streptococcus ( Strep ), Aeromonas ( Aerococcus ), the type species of the genus Aeromonas, officially named by WILLIAMS et al. in 1953. It is a bacterium between Staphylococcus and Streptococcus, but closer to the latter. Similar to Staphylococcus and Enterococcus in that it can grow in 6.5% sodium chloride broth, but it is catatase negative, small colony, α-hemolysis, etc. are closer to Streptococcus. [0003] The bacterium is an opportunistic pathogenic bacterium, which ...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K35/76A61P31/04A23K10/18A01N63/00A01P1/00C12R1/92
CPCA01N63/00A61K35/76A23K10/18A61P31/04C12N7/00C12N2795/10121C12N2795/10131C12N2795/10132
Inventor 韩文瑜顾敬敏袭恒豫程梦珺
Owner JILIN UNIV
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