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Wide-splitting-spectrum ultraviolet-resistant proteus mirabilis bacteriophage, composition thereof, kit and application of proteus mirabilis bacteriophage

A technology of Proteus mirabilis and phage, which is applied in the field of phage, can solve the problems of limited cracking range and UV resistance, and achieve the effects of no side effects, broad-spectrum bactericidal ability, and high proliferation efficiency

Pending Publication Date: 2022-06-03
PHAGELUX (NANJING) BIO-TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The above-mentioned prior art scheme has the following defects: the cracking range of the above-mentioned Proteus mirabilis phages is limited and not resistant to ultraviolet rays, and it is urgent to provide a wide cracking spectrum UV-resistant Proteus mirabilis phage

Method used

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  • Wide-splitting-spectrum ultraviolet-resistant proteus mirabilis bacteriophage, composition thereof, kit and application of proteus mirabilis bacteriophage
  • Wide-splitting-spectrum ultraviolet-resistant proteus mirabilis bacteriophage, composition thereof, kit and application of proteus mirabilis bacteriophage
  • Wide-splitting-spectrum ultraviolet-resistant proteus mirabilis bacteriophage, composition thereof, kit and application of proteus mirabilis bacteriophage

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Screening and purification of phage

[0042] The collected samples were centrifuged at 5000 r / min for 10 min and passed through a 0.22 μm filter.

[0043] (2) Phage enrichment for the target Proteus mirabilis in the sample The above filtrate was mixed with 2 times of LB liquid medium at a ratio of 1:1, inoculated with 100 μL of the target Proteus mirabilis strain, and enriched overnight.

[0044] (3) Phage screening and purification

[0045] Pick the above single plaque into 1 mL of SM buffer, shake at 150 rpm for 15 min, and dilute in a gradient manner, mix the dilution with 5 mL of LB semi-solid medium containing the target Proteus mirabilis, and pour it into the culture medium filled with LB solid medium. After the semi-solid medium solidified, it was incubated at 37°C overnight. This step was repeated 3 to 5 times to obtain a phage monoclonal sample, which was named Proteus mirabilis phage PMP1 (Proteus mirabilis phage PMP1). The form of Proteus mirabil...

Embodiment 2

[0047] Example 2 Determination of the optimal multiplicity of infection (MOI) of Proteus mirabilisphage PMP1 (Proteus mirabilisphage PMP1) on Proteus mirabilis

(1) Determination of Proteus mirabilis phage titer

Use SM solution as a diluent, and dilute the stock solution of Proteus mirabilis phage PMP1 (made by Example 1) to 10-fold gradient step by step. 8 times. Take l0 respectively 5 , l0 6 , l0 7 and l0 8 1000 μL of the diluted phage culture solution was evenly mixed with 300 μL of the host bacterial solution, and allowed to stand for 15 min to fully bind to the receptors on the bacterial surface. Add the above mixture into 5 mL of semi-solid agar medium cooled to 50°C, and immediately spread on the solidified solid agar plate after mixing. Three parallel samples should be made for each dilution, and the average of the three parallel samples of this dilution should be counted. Among them, the phage titer (PFU / mL) = the average number of plaques × the dilution facto...

Embodiment 3

[0049] Example 3 Deletion test of virulence gene or bad gene of Proteus mirabilis bacteriophage PMP1

[0050] Table 2 Main known virulence genes of lysogenic phages in pathogenic bacteria

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Abstract

The invention relates to the technical field of bacteriophages, in particular to a wide-splitting-spectrum ultraviolet-resistant proteus mirabilis bacteriophage and a composition thereof. The proteus mirabilis phage is a proteus mirabilis phage PMP1 (proteus mirabilis phage PMP1), and the proteus mirabilis phage is a proteus mirabilis phage. According to the proteus mirabilis phage PMP1, the sterilization rate of 105-106 PFU / mL of phage in a proteus mirabilis culture medium with the concentration of 102-103 PFU / mL on proteus mirabilis with the concentration reaches 97% or above, and the proteus mirabilis phage PMP1 has broad-spectrum sterilization capacity on the proteus mirabilis; the composition of the proteus mirabilis bacteriophage PMP1 can be used for cracking more proteus mirabilis, the cracking rate reaches 97% or above, and the proteus mirabilis bacteriophage PMP1 has higher cracking property.

Description

technical field [0001] The present application relates to the technical field of bacteriophage, in particular to a broad lysis spectrum UV-resistant Proteus mirabilis bacteriophage and its composition, kit and application. Background technique [0002] Proteobacteria are gram-negative saprophytic bacteria widely distributed in nature and belong to the family Enterobacteriaceae. The genus Proteus includes Proteous mirabilis, Proteous vulgaris, Proteous penneri, and Proteous myxofaciens. Domestic epidemiological data show that among the food-borne microbial pathogens, Salmonella, Vibrio parahaemolyticus and Proteus are the main representatives. In recent years, Proteus has accounted for a large proportion of bacterial food poisoning, especially the food poisoning caused by Proteus mirabilis accounts for 70% of Proteus food poisoning. [0003] The infection-related virulence factors of Proteus mirabilis are mainly manifested in the adhesion and invasiveness brought by fimbria...

Claims

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Application Information

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IPC IPC(8): C12N7/00A01N63/40A01N33/12A01P1/00A23K10/18A61K35/76A61P31/04A23B4/22C12R1/92
CPCC12N7/00A01N63/40A01N33/12A23K10/18A61K35/76A61P31/04A23B4/22C12N2795/10321C12N2795/10331C12N2795/10332Y02A50/30
Inventor 刘墨胡怿林徐旭凌黄杰费文斌陈海谢晓莉乔欢何四龙丛郁靳菊
Owner PHAGELUX (NANJING) BIO-TECH CO LTD
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