Detection method of phage

A detection method and phage technology, applied in the biological field, can solve the problems of reducing production costs, difficulty in grasping accuracy, and affecting the development of small and medium-sized enterprises, and achieve the effect of simple requirements, wide application range, and wide application range

Active Publication Date: 2015-01-28
山东京青农业科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] 1. There is a lack of effective detection methods for detecting bacteriophages in liquids such as sewers and decontamination tanks in fermentation production industrial plants
[0006] 2. The operation of the double-layer plate agar method in the laboratory is difficult, and its accuracy is difficult to grasp. It needs professional personnel to complete it. It is not suitable for large-scale and rapid detection in the fermentation industry.
[0007] 3. It is time-consuming and labor-intensive to detect bacteriophage with a bacterioplankton sampler, and a special bacteriophage sampler is required. The price is high, which is not conducive to reducing production costs, especially affecting the development of small and medium-sized enterprises.

Method used

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Examples

Experimental program
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Effect test

preparation example Construction

[0058] S100, preparation of culture medium

[0059] Prepare host medium suitable for the growth of phage host bacteria. During preparation, the medium preparation reagent is first sterilized, cooled to a suitable temperature, and then the medium preparation reagent is poured into a petri dish to prepare a host culture medium.

[0060] S200, coating of bacterial suspension

[0061] Spread the phage host bacterial suspension on the host medium prepared in step S100 to form a detection medium.

[0062] When coating, the bacterial lawn of the phage host bacteria can be inoculated in the sterilized culture medium in advance, mixed and fermented, and then coated on the host medium prepared by S100 to form a detection medium.

[0063] It should be noted that the phage host bacterium used in the present invention and the host medium suitable for its growth are not particularly limited, for example, the phage host bacterium can be selected from all available phage hosts, and the type...

Embodiment 1

[0072] Example 1 Detection of Bacillus subtilis phage in the air

[0073] Test site: Shandong Dongqing Technology Co., Ltd. fermentation plant area.

[0074] The detection method is as follows:

[0075] 1) Prepare nutrient agar medium:

[0076] The formula of nutrient agar medium is as follows:

[0077] 10.0 grams of peptone, 3.0 grams of beef extract, 5.0 grams of sodium chloride, 14.0 grams of agar, and 1000 milliliters of distilled water.

[0078] Autoclaved at 121°C for 15 minutes, cooled to 55°C and poured into a 9 cm diameter petri dish, after the culture medium cooled down, dried the medium in a drying oven at 45°C for 10 minutes to make a host Medium.

[0079] 2) Coating of bacterial suspension: The formula of the culture solution is: formula 10.0 grams of peptone, 3.0 grams of beef extract, 5.0 grams of sodium chloride, and 1000 milliliters of distilled water, autoclaved at 121 degrees Celsius for 15 minutes, and after cooling, Bacillus subtilis The bacterial law...

Embodiment 2

[0087] Example 2 Detection of Bacillus subtilis phage in liquid

[0088] Test site: Shandong Dongqing Technology Co., Ltd. fermentation plant area.

[0089] The detection method is as follows:

[0090] 1) Prepare nutrient agar medium:

[0091] The formula of nutrient agar medium is as follows:

[0092] 10.0 grams of peptone, 3.0 grams of beef extract, 5.0 grams of sodium chloride, 14.0 grams of agar, and 1000 milliliters of distilled water.

[0093] Autoclaved at 121°C for 15 minutes, cooled to 55°C and then poured into a 90 mm diameter petri dish. After cooling the culture medium, dry the medium in a drying oven at 45°C for 10 minutes to make a host Medium.

[0094]2) Coating of bacterial suspension: The formula of the culture solution is: formula 10.0 grams of peptone, 3.0 grams of beef extract, 5.0 grams of sodium chloride, and 1000 milliliters of distilled water, autoclaved at 121 degrees Celsius for 15 minutes, and after cooling, Bacillus subtilis Bacterial lawn is i...

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Abstract

The invention discloses a detection method of phage. The detection method comprises the following steps: S100, preparation of a culture medium: preparing a host culture medium which is suitable for growth of a phage host bacterium; S200, coating a bacterium suspension: coating a phage host bacterium suspension on the host culture medium to form a detection culture medium; S300, detection of the phage: detection of phage in gas: firstly, exposing and placing the detection culture medium in a to-be-detected region, then, culturing, and calculating the quantity of phages by using an OmeiLianski formula; and detection of the phage in a liquid: firstly, coating the detection culture medium with a to be detected liquid, then, culturing and calculating the quantity of the phages in the liquid. The detection method disclosed by the invention not only can be used for accurately detecting the quantity of the phages in gas of an industrial factory district, but also can be used for accurately detecting the quantity of the phages in liquids of the industrial factory district such as sewers and decontamination pools.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for detecting phages. Background technique [0002] Bacteriophage was independently discovered by Twort and D.Herelle in 1907 and 1909 respectively. Bacteriophage is a general term for viruses that infect microorganisms such as bacteria, fungi, actinomycetes or spirochetes. Some of them can cause the lysis of host bacteria. Hence the name bacteriophage. As a kind of virus, bacteriophage has some characteristics unique to viruses: the individual is small; it does not have a complete cell structure; it only contains a single nucleic acid. The phage genome contains many genes, but all known phages use bacterial ribosomes, various factors required for protein synthesis, various amino acids and energy production systems in bacterial cells to achieve their own growth and proliferation , once outside the host cell, the phage can neither grow nor replicate. [0003] The fermentat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/06C12R1/92
CPCC12Q1/06C12Q1/70
Inventor 郭中信冯阿芳赵丽郭倩
Owner 山东京青农业科技有限公司
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