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Administration method for protecting bacteriophage activity and application thereof

A drug delivery method and phage technology, which are applied in the field of biomedicine and can solve problems such as no phage drug delivery method yet.

Inactive Publication Date: 2012-03-14
万学勤
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no drug delivery method that uses live cells of phage host bacteria to protect phages.

Method used

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  • Administration method for protecting bacteriophage activity and application thereof
  • Administration method for protecting bacteriophage activity and application thereof
  • Administration method for protecting bacteriophage activity and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1. Isolation of bacteriophage (1) Preparation of the target bacterial suspension: Take the specimen of the infected site for isolation and pure culture as a target bacterial slant, add sterile water to wash the bacterial moss, and prepare a bacterial suspension. The medium and culture method used are all based on the culture of the target bacteria (the same below).

[0021] (2) Proliferation and culture of phages in the sample: In a three-fold concentrated liquid medium, filter-sterilized sewage samples were added in 1:2, and the target bacteria suspension was inoculated, so that the phages in the samples were cultured together with the target bacteria. The specific culture time is determined according to different bacteria, such as Escherichia coli and other bacteria culture for 12-24h; Helicobacter pylori culture for 5-7d (the same below).

[0022] (3) Preparation of phage lysate: after centrifuging the above mixed culture solution at 2500 r / min for 15 min, th...

Embodiment 2

[0024] Example 2. Purification of bacteriophage (1) After the existence of bacteriophage was confirmed, a phage double-layer plate quantitative purification test was performed. The first tube uses an inoculation loop to take 2 loops of filtrate and add 0.2ml of suspension, the second tube of filtrate 5 loops add 0.2ml of bacterial suspension, the third tube of filtrate 10 loops add 0.2ml of bacterial suspension, and the fourth tube is made with 0.2ml of filtrate alone. Filtrate control, the fifth tube used 0.2ml bacterial suspension alone as bacterial control. Mix the tubes evenly and stand at 37°C for 15-30min.

[0025] (2) Take the upper layer of 0.8% agar medium, dissolve and cool to 48°C, add 4 ml to 0.2 ml of the above mixture of phage and bacteria, and mix immediately.

[0026] (3) and immediately pour it onto the bottom medium and spread it evenly. Incubate at 37°C for a certain period of time.

[0027] (4) The single plaques grown at this time are often inconsistent...

Embodiment 3

[0030] Example 3. Preparation of high-titer bacteriophage The phage obtained just after isolation and purification is often not high in titer and needs to be propagated. Mix the purified phage filtrate with the liquid medium at a ratio of 1:10, then add an appropriate amount of host bacterial suspension (it can be equal to or 1 / 2 of the amount of the phage filtrate), cultivate and multiply, and repeat the number of transplants. second, and finally filtered to obtain a high-titer phage product.

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PUM

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Abstract

The invention discloses an administration method for protecting bacteriophage activity and application thereof. The method utilizes host bacteria of the bacteriophage to protect the bacteriophage, thereby enhancing resistance of the bacteriophage. The method aims to help the bacteriophage to break body immunization barriers, so that the bacteriophage can complete a first propagation under protection of the host bacteria and release numerous filial generation bacteriophages and further singularly invade and crack infectious bacteria. The method of the invention can be applied to bacteriophage therapies of infections of all parts and organs of body caused by prokaryotic and eukaryotic cell microbes. The invention reduces interference of body resistance on the bacteriophage therapy, amplifies and magnifies treatment dosage and treatment effect of the bacteriophage, and provides a new, practical and effective administration method for enforcing clinical bacteriophage treatment on microbe infectious diseases.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a drug administration method for protecting bacteriophage activity and its application. Background technique [0002] Phages are viruses that infect microorganisms such as bacteria, fungi, actinomycetes, and spirochetes, and are highly specific to host bacteria. According to the status and results of their infection of the host bacteria, phages are divided into two types, one of which can replicate and proliferate in the host bacteria cells, produce many progeny phages, and finally lyse the bacteria, called potent or toxic phages. After infecting bacteria, virulent phages can proliferate in bacteria and kill bacteria without toxicity to animals and plants. Therefore, virulent phages are expected to be better antibacterial drugs due to their unique natural characteristics. [0003] As early as the 1930s, there were reports of successful treatment of cholera with phages. So f...

Claims

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Application Information

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IPC IPC(8): A61K35/76A61P31/04
Inventor 万学勤李宏鸣崔志新唐冬生
Owner 万学勤
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