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A high-temperature-resistant strong Proteus phage rdp-sa-20018 and its application

A technology of RDP-SA-20018 and Proteus, which is applied in the direction of bacteriophage, virus/phage, medical raw materials derived from virus/phage, etc., can solve the problems of drug resistance of pathogenic bacteria, untimely treatment, ineffective medication, etc., to achieve High potency, wide cracking spectrum, good control effect

Active Publication Date: 2022-03-18
RECOM QINGDAO BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a strain of high temperature resistant Proteus speciesphage (Proteus speciesphage) RDP-SA-20018. lead to problems such as drug ineffectiveness

Method used

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  • A high-temperature-resistant strong Proteus phage rdp-sa-20018 and its application
  • A high-temperature-resistant strong Proteus phage rdp-sa-20018 and its application
  • A high-temperature-resistant strong Proteus phage rdp-sa-20018 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Isolation and purification, drug resistance and virulence gene identification of pathogenic Proteus BS-20013

[0030] (1) Isolation and identification of Proteus

[0031] Samples were taken from the viscera of sick and dead chickens in a farm in Liaoning Province, and the aseptic method was used to draw a line on the SS medium. After culturing at 37°C for 18-24 hours, there were white neat colonies with a black center and a white edge on the SS agar plate. About 2-3 mm; pick typical colonies and streak them on LB agar plate, after culturing at 37°C for 10-24 hours, they will show diffuse growth on LB agar plate, pick typical colonies and continue to streak and purify for 3 times, then pick a single colony Inoculate in 5 mL of LB liquid medium, culture at 37°C with shaking at 200 rpm for 8 hours, and obtain a uniform turbid bacterial suspension. Through 16sRNA molecular identification, it was determined to be Proteus, and one of them was named BS-20013.

[00...

Embodiment 2

[0040] Example 2 Isolation and identification of Proteus phage RDP-SA-20018

[0041] (1) Sampling: The sample used in the test of the present invention was collected from the sewage around the diseased farm in Liaoning Province in 2020 as a water sample isolated from bacteriophage.

[0042] Take 10 ml of sewage, centrifuge at 10,000rpm for 5 minutes, remove larger impurities and most of the bacteria, and then use a 0.22 micron microporous membrane to filter and sterilize. Take 3 milliliters of the filtrate and 3 milliliters of the host bacterial suspension, add them together into 20 milliliters of autoclaved 2216E liquid medium, and then place them in a 37°C incubator for overnight culture. After culturing, take 5 ml, centrifuge at 10,000 rpm for 5 minutes, and then filter and sterilize with a 0.22 micron microporous filter. The filtrate is the stock solution intended to contain phage. Then, use the double-plate method to identify whether there are phages. If there are phage ...

Embodiment 3

[0047] Morphological observation of embodiment 3 phage

[0048] Using phosphotungstic acid negative staining method: drop phage liquid (titer 10 10 pfu / m L) 100 μL, place the membrane side of the copper mesh on the phage droplet, take it off after 10 minutes, and let it dry naturally in the air for 2-3 minutes. Then drop a drop of 2% phosphotungstic acid (PTA) aqueous solution on the copper grid for staining, take it off after 10 minutes, dry it in the air for 10-15 minutes, observe with an electron microscope, and select a clear phage image to take pictures.

[0049] Depend on figure 1 It can be seen that the head of the phage is regular hexagonal, the diameter of the head is about 52nm, and the length of the tail is 110nm, which is a long-tailed phage.

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Abstract

The invention discloses a high-temperature-resistant potent proteus phage RDP‑SA‑20018, the preservation number is CGMCC No.21405, the full length of the gene is 42626bp, and it is found after sequencing the whole gene that it does not have virulence genes and lysogenic genes, The Proteus phage RDP‑SA‑20018 provided by the present invention has higher temperature tolerance and wider acid-base tolerance range, which is conducive to industrial production and preparation; it is found by lysis rate experiment and RTD experiment that the lysis of the bacteriophage The rate is 98%, and it still has a good cracking effect on pathogenic bacteria. The phage has the characteristics of excellent performance and safety. The phage can be used in the preparation of medicines for preventing and or treating diseases caused by proteus.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a high-temperature-resistant strong Proteus phage RDP-SA-20018 and its application in pathogenic Proteus infection of poultry animals. Background technique [0002] Proteus is a common opportunistic pathogen that is widely distributed in nature and can infect humans and various livestock and poultry, and cause body death. In recent years, the prevalence of Proteus has continued to expand, and with the non-standard use of antibiotics, the drug resistance of the bacteria has continued to increase, which has brought huge economic losses to human and animal husbandry. [0003] Phage is a bacterial virus with high abundance in nature, strong host specificity, and will not damage the micro-ecological environment. After phages infect host bacteria, new phages will be released during lysis, and a small amount of phages can have a good control effect. Contents of the invention [0004] Th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00A01N63/40C11D3/38C11D3/48A61K35/76A61K45/06A61P31/04A01P1/00A23K10/18A23K50/75
CPCC12N7/00A01N63/40C11D3/381C11D3/48A61K35/76A61K45/06A61P31/04A23K10/18A23K50/75C12N2795/10311C12N2795/10331C12N2795/10332
Inventor 杜新永李先胜马如霞罗成盛赵丹丹王立坤张庆刘玉庆盖春云刘爽
Owner RECOM QINGDAO BIOTECH CO LTD
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