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Carrier for separating bacteriophage and preparation method and application thereof

A phage and carrier technology, applied in the field of virus isolation, can solve the problem of low success rate of phages, and achieve the effect of increasing the probability of encounter and adsorption

Active Publication Date: 2017-06-09
JIMEI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, using the traditional phage isolation method, when the phage content is less than 10 2 / m 3 The success rate of phage isolation from sewage, sediment, seawater, etc. is very low

Method used

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Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0023] The present invention also provides the preparation method of the carrier for separating phage described in the above technical scheme, comprising the following steps:

[0024] 1) inoculating Vibrio parahaemolyticus in the culture medium of the host bacteria to obtain the culture medium of the host bacteria;

[0025] 2) Submerge the ceramic carrier in the host bacteria culture solution obtained in step 1), and dry the soaked ceramic carrier to a water content of 45-55% to obtain a carrier for isolating bacteriophage.

[0026] In the present invention, Vibrio parahaemolyticus is inoculated in the culture medium of the host bacteria and cultivated to obtain the culture solution of the host bacteria. In the present invention, the host bacteria culture medium preferably includes the following components: beef extract 1-8g / L, peptone 5-15g / L, sodium chloride 15-55g / L, and the balance is water or yeast extract Cream 1-8g / L, tryptone 5-15g / L, sodium chloride 20-60g / L, the bal...

Embodiment 1

[0049] Under sterile conditions, Vibrio parahaemolyticus was inoculated in the host bacteria culture medium and cultured at a temperature of 28°C and a rotation speed of 150rpm for 14 hours to obtain a culture medium of the host bacteria. In the culture medium, parahaemolyticus Vibrio content in 10 8 More than cfu / ml; the host bacteria culture medium is: beef extract 5g / L, peptone 10g / L, sodium chloride 30g / L, and the balance is water.

[0050] Soak the ceramic carrier in the host bacterium culture solution for 2 hours, the mass ratio of the ceramic carrier to the host bacterium culture solution is 1g:1ml, take it out after soaking, and dry it under ventilated conditions so that the water content of the ceramic carrier is 50%, repeat the steps of soaking and drying 3 times, and then wrap tightly with 40 mesh gauze to obtain the carrier for isolating bacteriophage.

[0051] Put the carrier for isolated phage into the flowing seawater for 4 hours at a depth of 50 cm, and then p...

Embodiment 2

[0053] Under aseptic conditions, Vibrio parahaemolyticus was inoculated in the culture medium of the host bacteria and cultivated for 24 hours at a temperature of 35°C and a rotation speed of 200rpm to obtain a culture medium of the host bacteria, in which the parahaemolyticus Vibrio content in 10 8 More than cfu / ml; the host bacteria culture medium is: beef extract 5g / L, peptone 10g / L, sodium chloride 30g / L, and the balance is water.

[0054] Soak the ceramic carrier in the host bacterium culture solution for 1 hour, the mass ratio of the ceramic carrier to the host bacterium culture solution is 0.5g:1.5ml, take it out after soaking, and dry it under ventilated conditions, so that the water content of the ceramic carrier The yield was 40%, the steps of soaking and drying were repeated 3 times, and then tightly wrapped with 54-mesh gauze to obtain the carrier for isolating phage.

[0055] Put the carrier for isolated phage into the flowing seawater for 6 hours at a depth of 8...

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PUM

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Abstract

The invention provides a carrier for separating bacteriophage. The carrier for separating the bacteriophage comprises a ceramic carrier body, and vibrio parahaemolyticus which is loaded on the ceramic carrier body. With the adoption of the carrier, bacteriophage of vibrio parahaemolyticus in seawater, fresh water and wastewater can be quickly separated, and the separating efficiency reaches 80%; the identifying result shows that the separated bacteriophage is vibrio parahaemolyticus bacteriophage.

Description

technical field [0001] The invention belongs to the technical field of virus isolation, and in particular relates to a vector for isolating bacteriophage, a preparation method and application thereof. Background technique [0002] In recent years, due to the high-density farming in aquaculture and the widespread use of antibiotics, a large number of drug-resistant strains have emerged. As a more effective treatment, bacteriophages are attracting more and more attention. Therefore, rapid phage isolation is the first step in conducting phage therapy research. Phages widely exist in the natural environment, where there are bacteria, there may be corresponding phages. For example, in human and animal excrement or polluted well water and river water, there are often phages of intestinal bacteria. Phages also widely exist in the marine environment, and bacteria, cyanobacteria, and eukaryotic microorganisms in the ocean have all found their corresponding phages. [0003] At pres...

Claims

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Application Information

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IPC IPC(8): C12N11/14C12N7/00C12R1/63
CPCC12N7/00C12N11/14C12N2795/00051
Inventor 林茂付汉清
Owner JIMEI UNIV
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