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A bacteriophage str-pap-1 and a composition for preventing and treating Streptococcus parauberis infection using the phage as an active ingredient

An active ingredient and phage technology, applied in the direction of phage, virus/phage, medical raw materials derived from virus/phage, etc., to achieve the effect of small side effects and high specificity

Active Publication Date: 2018-03-20
INTRON BIOTECHNOLOGY INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, after Flemming discovered penicillin, antibiotics were widely used, so research on phages was limited to some Eastern European countries and the former Soviet Union.
However, after 2000, due to the increase of antibiotic-resistant bacteria, the existing antibiotics showed certain limitations

Method used

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  • A bacteriophage str-pap-1 and a composition for preventing and treating Streptococcus parauberis infection using the phage as an active ingredient
  • A bacteriophage str-pap-1 and a composition for preventing and treating Streptococcus parauberis infection using the phage as an active ingredient
  • A bacteriophage str-pap-1 and a composition for preventing and treating Streptococcus parauberis infection using the phage as an active ingredient

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Embodiment 1: isolate the phage that can kill Streptococcus parauberis

[0037] The samples obtained from the natural environment or animal samples were used in the screening of phages capable of killing Streptococcus parauberis. In addition, the Streptococcus parauberis used for isolation of the phage was previously isolated and identified as Streptococcus parauberis by the present inventors.

[0038] Next, the phage isolation process is described in detail. The collected samples were added together with Streptococcus parauberis to TSB (Tryptic Soy Broth) medium inoculated at a ratio of 1 / 1000 (casein digestion medium, 17g / L; soybean digestion medium, 3g / L; right Glucose, 2.5g / L; NaCl, 5g / L; Dipotassium hydrogen phosphate, 2.5g / L), and then cultured overnight at 30°C with shaking. After culturing, centrifugation was performed at 8,000 rpm for 20 minutes to recover the supernatant, which was then filtered through a 0.45 μm filter. The presence or absence of bacteri...

Embodiment 2

[0044] Embodiment 2: Genome isolation and sequence analysis of bacteriophage Str-PAP-1

[0045] The genome of phage Str-PAP-1 was isolated as follows. For genome isolation, the phage suspension obtained by the same method as in Example 1 was used. First, in order to remove DNA and RNA of Streptococcus parauberis that may be contained in the suspension, 200 U of DNase I and RNase A were respectively added to 10 ml of the phage suspension, and then left at 37° C. for 30 minutes. After standing for 30 minutes, in order to remove the activity of DNase I and RNase A, 500 μl of 0.5M ethylenediaminetetraacetic acid (EDTA) was added and then left to stand for 10 minutes. Then, it was left to stand at 65° C. for an additional 10 minutes. In order to disintegrate the outer wall of the phage, 100 μl of proteinase K (20 ㎎ / ml) was added and reacted at 37° C. for 20 minutes. Then, 500 µl of 10% sodium dodecyl sulfate (SDS) was added, and the mixture was reacted at 65° C. for 1 hour. A...

Embodiment 3

[0049] Embodiment 3: the investigation to the ability of bacteriophage Str-PAP-1 killing Streptococcus parauberis

[0050] The ability of the isolated phage Str-PAP-1 to kill Streptococcus parauberis was investigated. The same method as in Example 1 was used for the investigation of the killing ability, and it was observed whether a transparent ring was formed by a drop test. The Streptococcus parauberis used in the killing ability analysis was isolated and identified by the present inventors as Streptococcus parauberis, and there are 55 species in total. Phage Str-PAP-1 has the ability to kill 35 strains of Streptococcus parauberis used in the experiment. Representative experimental results such as figure 2 shown. In addition, the effects of phage Str-PAP-1 on Edwardsiella tarda, Vibrio anguillarum, Vibrio ichthyoenteri, Lactococcus garvieae and Streptococcus dolphin were investigated. iniae) killing ability. The results showed that phage Str-PAP-1 had no ability to k...

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Abstract

The present invention relates to a composition comprising a bacteriophage Str-PAP-1, a bacteriophage that can kill Streptococcus parauberis after being infected with Streptococcus parauberis isolated from nature, as an active ingredient, and the use of the composition to prevent and treat Methods of Streptococcus parauberis infection. The phage Str-PAP-1, which is an active ingredient of the composition according to the present invention, is characterized in that it has the genome represented by SEQ ID NO: 1 capable of killing Streptococcus parauberis.

Description

technical field [0001] The present invention relates to a composition which can be used to prevent or treat the infection of Streptococcus parauberis, which uses the bacteriophage isolated from nature and capable of killing Streptococcus parauberis after being infected with Streptococcus para And the method of treating Streptococcus parauberis infection. Specifically, it relates to a composition for preventing and treating Streptococcus parauberis infection and after infection, the composition will have the genome represented by sequence number 1 capable of specifically killing Streptococcus parauberis The isolated bacteriophage contains said phage as an active ingredient; and the method of preventing Streptococcus parauberis infection and post-infection treatment using the composition. Background technique [0002] At present, the proportion of aquaculture in the entire aquatic industry continues to increase. From a global perspective, the amount of fish caught has stagna...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00A61K35/76A61P31/04A23K10/18C12R1/92A23K20/195
CPCA23K10/18A23K20/195A23K50/80A61K35/76A61P31/04C12N7/00C12N2795/10321C12N2795/10332
Inventor 尹成俊全秀娟权安成许荣宰金正美姜尚铉
Owner INTRON BIOTECHNOLOGY INC
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