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Device and method for high-throughput screening of active strains with potential biological anti-corrosion function

A high-throughput, strain-based technology, applied in the field of microbial detection, can solve problems affecting the detection results of the antibacterial activity of actinomycetes

Inactive Publication Date: 2021-03-26
GUANGDONG INST OF MICROBIOLOGY GUANGDONG DETECTION CENT OF MICROBIOLOGY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

attached figure 2 , the actinomycete strains screened out on the selective culture plate produce red, blue and yellow pigment substances respectively in the later stage of culture, thus affecting the test results of the antibacterial activity of actinomycetes

Method used

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  • Device and method for high-throughput screening of active strains with potential biological anti-corrosion function
  • Device and method for high-throughput screening of active strains with potential biological anti-corrosion function
  • Device and method for high-throughput screening of active strains with potential biological anti-corrosion function

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] This embodiment introduces in detail the process of using the high-throughput screening device of the present invention to screen the bacterial strains with antibacterial activity, specifically as follows:

[0036] Step 1: Obtain the pure culture fermentation supernatant of different strains to be tested;

[0037]Step 2: Pour the unsolidified agar medium containing the indicator bacteria into the transparent vessel 1 of the device, and wait for it to solidify;

[0038] Alternatively, step 2: Pour the unsolidified agar medium into the transparent vessel 1 of the device, and after it solidifies, apply a certain concentration of indicator bacteria;

[0039] Step 3: Place the porous cell culture plate 2 in the transparent vessel 1. At this time, the bottom 3 is in contact with the surface of the solid medium 4; absorb 20-50 μl of fermentation supernatants of different strains and add them to the porous cells of the device respectively each bottom 3 of the culture plate 2; ...

Embodiment 2

[0041] Example 2 High-throughput screening of actinomycete strains with bacteriostatic activity against Escherichia coli

[0042] Collect soil samples from Baiyun Mountain in Guangzhou, weigh 10g of soil samples, mix them in 100ml of sterile water, and serially dilute 10 3 、10 4 、10 5 times. Spread the dilution of the soil sample on the selection plate of actinomycetes for culture, and use a sterile inoculation loop to pick up multiple suspected actinomycete single colonies that grow well on the selection solid medium; No. 1 liquid culture medium in a 50mL sterile centrifuge tube; place the inoculated strain in a shaker at 28°C and 160r / min for about 7 days; place the fermentation broth of the cultured strain in a 50mL centrifuge tube in a centrifuge Centrifuge at 8000r / min in the machine to obtain the fermentation supernatant of the strains; use a pipette to absorb 20-50 μl of the fermentation supernatants of different strains and add them to the high-throughput screening ...

Embodiment 3

[0047] Example 3 High-throughput screening of actinomycete strains with bacteriostatic activity against Aspergillus niger

[0048] Collect soil samples from Baiyun Mountain in Guangzhou, weigh 10g of soil samples, mix them in 100ml of sterile water, and serially dilute 10 3 、10 4 、10 5 times. Spread the dilution of the soil sample on the selection plate of actinomycetes for culture, pick a single colony, and follow the Figure 5 The process steps for liquid culture, centrifuge to take the fermentation supernatant, pour 20ml of PDA medium at the bottom of the quartz vessel, and after it solidifies, the concentration of the bacteria to be coated is 10 5-6 200 μl of cfu / ml Aspergillus niger liquid, placed in a 96-hole plate, and 50 μl of fermentation supernatant of different strains to be screened was added to each well plate, covered with the upper cover of the quartz vessel, and placed in a culture medium at 37°C After overnight incubation, the activity of the screened stra...

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Abstract

The invention provides a device for high-throughput screening of bacteriostatic active strains. The device comprises a transparent vessel (1) and a porous cell culture plate (2), culture deep holes are uniformly distributed in the cell culture plate, bottoms (3) of the culture deep holes are hollow, the porous cell culture plate (2) is placed in the transparent vessel (1), a distance is reserved between the bottom (3) of the cell culture plate and the bottom of the inner wall of the transparent vessel, and the bottom of the cell culture plate does not make contact with the bottom of the innerwall of the transparent vessel; the transparent vessel is used for containing a solid culture medium (4) containing indicator bacteria, and the porous cell culture plate is used for containing a culture supernatant of to-be-detected strains; and when the transparent vessel contains an agar culture medium containing the indicator bacteria, the bottom (3) of the porous cell culture plate is in contact with the surface of the solid culture medium (4). The porous cell culture plate in the device can be used for detecting the bacteriostatic activity of the multiple strains at the same time in a high-throughput mode, and is not limited to the number of holes of the cell culture plate, of the multiple culture holes can be prepared according to actual requirements, and each hole is used for detecting one to-be-detected strain. The indicator bacteria can further be flexibly replaced, and the antibacterial spectrum of the to-be-detected bacteria can be rapidly detected.

Description

technical field [0001] The invention relates to the field of microbial detection, in particular to a device and method for high-throughput screening of active strains with potential biological antiseptic functions. Background technique [0002] With the development of society, food safety has become one of the key issues of today's society. In the process of food processing, transportation and storage, it is inevitable to breed various microorganisms. Spoilage fungi such as Aspergillus flavus, Aspergillus niger and Penicillium citrinum and pathogens such as Escherichia coli, Staphylococcus aureus and Listeria Bacteria can have harmful effects on human health. The development of efficient, safe and healthy natural preservatives to replace the traditional chemically synthesized preservatives (such as sodium benzoate and potassium sorbate) widely added in food today is the key direction of food industry research in various countries in the world today. Among them, microbial p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/34C12Q1/04
CPCC12M41/36C12Q1/04
Inventor 吴清平何莹龙庞锐张菊梅莫树平柏建玲丁郁王涓薛亮陈谋通谢新强
Owner GUANGDONG INST OF MICROBIOLOGY GUANGDONG DETECTION CENT OF MICROBIOLOGY
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