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Drug-resistant coli phage and application thereof

A technology of Escherichia coli and bacteriophage, applied in the field of microbial medical engineering, can solve the problems of increasing the number of Escherichia coli, increasing the incidence rate, etc., and achieve the effect of good killing effect

Active Publication Date: 2019-01-22
NANJING MEDICAL UNIV
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Problems solved by technology

In addition, long-term use of antibiotics will increase the number of multi-drug-resistant E. coli carried in feces, resulting in a 10-fold increase in the incidence of urinary tract infections in women

Method used

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  • Drug-resistant coli phage and application thereof
  • Drug-resistant coli phage and application thereof
  • Drug-resistant coli phage and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0018] 1. Preliminary isolation and purification of phage

[0019] Sewage sample collection and processing:

[0020] Escherichia coli MG1655 was used to isolate and enrich virulent phages in Nanjing wastewater. Sewage samples were taken from polluted rivers in Nanjing City, and the sewage samples were filtered and sterilized with a 0.45 μm filter. Take 1 mL of the sewage filtrate and 500 μL of the MG1655 overnight culture solution and place it in 5 mL of liquid LB medium, shake and culture at 200 rpm at 37°C for 24 hours to enrich the phage. Take 1 mL of culture solution, centrifuge at 14,000 g for 10 minutes, and filter the supernatant with a 0.45 μm filter membrane. Take 10 μL of filtrate, add 100 μL of overnight bacterial solution (MG1655), then add 3.5 mL of upper agar (semi-solid LB medium) and mix well, spread on LB agar plate (double-layer agar plate method), observe phage plaque after overnight culture form. The phages isolated for the first time are different in s...

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Abstract

The invention discloses drug-resistant coli phage and application thereof. The classification of the drug-resistant coli phage is named as Escherichia phage vB_EcoP-EG1, the reservation number is CCTCC M 2018562, and the preservation date is August 21, 2018. The phage Escherichia phage vB_EcoP-EG1 has stable properties, a short life cycle and high lysis ability to host bacteria. The results of a genome and a protein map show that the phage Escherichia phage vB_EcoP-EG1 does not encode toxic proteins. At the same time, the phage Escherichia phage vB_EcoP-EG1 can infect and lyse a plurality of strains of clinically and randomly isolated multi-drug resistant escherichia coli, and particularly has a good killing effect on uropathogenic escherichia coli. In conclusion, Escherichia phage vB_EcoP-EG1 is a good candidate phage for phage therapy, can be used as a single preparation or mixed with other phages to form a cocktail preparation for fighting against infection caused by the clinical multi-drug resistant escherichia coli.

Description

technical field [0001] The invention belongs to the field of microbiological medical engineering, and in particular relates to a phage capable of lysing clinically isolated multi-strain drug-resistant colibacillus and its application. Background technique [0002] Virulent phages can infect and lyse bacteria and have been used to kill pathogenic bacteria at the beginning of the last century. Subsequently, due to the discovery and large-scale application of cheap and effective antibiotics, they were gradually ignored. Only a few Eastern European countries (such as Georgia, Russia, etc.) have been insisting on using phage therapy. In recent years, due to the continuous emergence of drug-resistant bacteria, phages have once again attracted people's attention. Especially in the past decade or so, a large number of new phages targeting drug-resistant bacteria have been discovered and widely studied. Many in vitro and in vivo experiments have proved that phages can kill a variety...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K35/76A61P31/04
CPCA61K35/76A61P31/04C12N7/00C12N2795/10221Y02A50/30
Inventor 刘晓秋古雨于鑫焱刘根焰张学森
Owner NANJING MEDICAL UNIV
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