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Shigella dysenteriae phage SSE1 and application thereof

A Shigella dysenteriae and phage technology, applied in phage, virus/phage, applications, etc., can solve problems such as restrictions on the use of antibiotics, rapid superbacterial infection, and human health hazards, achieving rapid and efficient lysis, improving universality and Effectiveness, the effect of expanding the host spectrum

Inactive Publication Date: 2020-05-01
UNIVERSITY OF CHINESE ACADEMY OF SCIENCES
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Benefits of technology

The technology described by this patented solution allows us to create an antibiosis agent that targets specific types of germs called shiga bacteraemia type B (SSB) without causing harmful side reactions like other organisms such as Escherichia coli K-12a strain). This makes it possible to use these agents effectively against various diseases caused by those microorganism populations.

Problems solved by technology

The technical problem addressed by this patents relates to finding ways to treat shigae disease without relying on chemical or biological agents like chloramphenicol (CIP) due to concerns over safety issues associated with these methods used against other types of microorganisms such as Salmonellus spp., Escherichia coli O157. Additionally, current approaches require long periods of time before they work out but may be effective only if certain conditions exist during the course of illnesses preventing them from spreading even more rapidly.

Method used

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  • Shigella dysenteriae phage SSE1 and application thereof
  • Shigella dysenteriae phage SSE1 and application thereof
  • Shigella dysenteriae phage SSE1 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] The separation and purification of embodiment 1 phage

[0020] The sewage sample used in the experiment of the present invention was collected in September 2018 in an aeration tank of a sewage treatment plant in Beijing, and used as a water sample for separating bacteriophage.

[0021] Centrifuge the sewage sample for 15min, take 25mL of sewage sterilized by filtration with a 0.22μm microporous membrane; add CaCl 2 Mother liquor to a final concentration of 1mmol / L, let it stand; add 20mL NB liquid medium, then add 2.0mL of logarithmic phase host bacterial suspension in a 50ml centrifuge tube, mix well; after standing for 15min, shake overnight at 37°C (10h ). The next day, add 2 mL of chloroform, centrifuge at 8,000 g for 30 min at 4°C; take 15 mL of supernatant, add 20 mL of NB culture medium and 1.0 mL of logarithmic phase host bacterial suspension, shake well, and place at room temperature for 30 min; shake at 37°C for 120 r min -1 Cultivate for 3-6 hours, take it ...

Embodiment 2

[0023] Morphological observation of embodiment 2 bacteriophage

[0024] Take 20 μL of the phage suspension and drop it on the copper grid, wait for its natural precipitation for 10 minutes, blot it dry from the side with dry filter paper, let it air for about 1 minute, add 1 drop of 1% uranyl acetate on the copper grid, stain for 2 minutes, and then use it carefully to dry Blot the excess dye from the side of the filter paper, let it dry naturally in the dark for 30 minutes, and observe it with a transmission electron microscope (JEM-1400).

[0025] The results of transmission electron microscopy are shown in figure 2 , the results show that the phage SSE1 has an icosahedral head and a retractable tail, the head length is about 123.5nm, the head diameter is about 82nm, the long tail is about 130nm long, and the tail diameter is about 25nm, according to the International Virus Taxonomy Organization ( According to the eighth report of ICTV virus taxonomy, the phage SSE1 strain...

Embodiment 3

[0026] Example 3 Phage Genome Analysis and Identification

[0027] Phage whole genome sequencing and analysis: Illumina NextSeq500 was used to perform PE2×150 DNA sequencing of the samples. Then, the optimized sequence was spliced ​​using spades v.3.11.1 splicing software, and the optimal assembly result was obtained. Use the biological software PHASTER to predict and analyze the open reading frame of the genome, and use NCBI Blastp to complete the preliminary annotation of functional genes, and use tRNAscan-SE (http: / / lowelab.ucsc.edu / / tRNAscan-SE / ) to predict tRNA online, Use CGView Server software (http: / / cgview.ca / ) to complete the drawing of the whole genome circle map (see image 3 ), a phylogenetic tree was constructed using MEGA X software based on the whole genome data.

[0028] Whole-genome analysis shows that the genome of phage SSE1 is a circular double-stranded structure with a full length of 169744bp and a G+C% content of 37.51%. There are 2 predicted tRNAs and...

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Abstract

The invention discloses a Shigella dysenteriae phage SSE1 and application thereof, and belongs to the field of biotechnology. The disclosed Shigella dysenteriae phage SSE1 has the collection number CGMCC No. 18853, provides a selectable basic material for the prevention and control of pathogens, and enriches the phage library; through the isolation and identification of phage SSE1 and the whole genome sequencing, functional genes and proteins are further mined, one or more enzymes having a lytic effect on pathogenic bacteria are provided for phage treatment and biocontrol so as to expand the host spectrum and improve the wide and effective application of phages; and the phage SSE1 can quickly and efficiently lyse dysentery bacillus, can be applied to pollution control of dysentery bacillusin sewage treatment systems and removal of drug-resistant dysentery bacillus.

Description

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Claims

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Application Information

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Owner UNIVERSITY OF CHINESE ACADEMY OF SCIENCES
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