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Method for quickly separating Geobacillus stearothermophilus spores from nutrient thalli

A technology of Geobacillus and thermophilic fat, which is applied in the field of microbial spore fermentation, can solve the problems of spore loss, poor spore and cell effect, and difficulty in distinguishing, and achieves the effect of thorough separation and good quality control.

Inactive Publication Date: 2020-07-17
NANJING JUSHA DISPLAY TECH +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the key technical issue of how to selectively suspend bacterial cells has not been clarified. The bacterial cells and spores can only be selected under a microscope, which is difficult to distinguish with the naked eye. Reduced; the actual operation is cumbersome, and the effect of separation of spores and bacteria is not good

Method used

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  • Method for quickly separating Geobacillus stearothermophilus spores from nutrient thalli
  • Method for quickly separating Geobacillus stearothermophilus spores from nutrient thalli

Examples

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specific Embodiment 1

[0025] The present invention provides a kind of method for rapidly separating Geobacillus stearothermophilus spores and vegetative thalline, comprising the following steps:

[0026] In step one, the suspension containing spores and bacterial cells is collected in a sterile centrifuge container.

[0027] Step 2, put the centrifuge container into the centrifuge equipment for centrifugation. In the prior art, the centrifugal separation speed of Geobacillus stearothermophilus spores is generally 4000-7500 rpm, and the centrifugation time is 30-60 minutes. In the present invention, in order to realize the sedimentation of the spores and the suspension of the vegetative bacteria, it is necessary to strictly control the rotation speed at 7500-8500 rpm and the centrifugation time for 10-15 minutes. In this embodiment, the rotation speed is preferably 8000 rpm, the centrifugation temperature is 4° C., and the centrifugation time is 12 minutes, which can realize the suspension of veget...

specific Embodiment 2

[0033] The invention provides a kind of method for rapidly separating Geobacillus stearothermophilus spores and vegetative thalline, comprising the following steps:

[0034] In step one, the suspension containing spores and bacterial cells is collected in a sterile centrifuge container.

[0035] Step 2, put the centrifuge container into the centrifuge equipment for centrifugation, the centrifugation speed is selected as 7500-8500rpm, preferably 7500rpm in this embodiment, the centrifugation temperature is 4°C, and the centrifugation time is 15min, which can realize the suspension of vegetative bacteria and the separation of spores precipitation.

[0036] Step 3: Discard the upper liquid of the suspension after centrifugation, add an appropriate amount of suspending medium, and resuspend the sediment layer in the suspending medium. The suspending medium includes sterile purified water, suspending agent solution or density gradient separation liquid. This implementation In the ...

specific Embodiment 3

[0041] The invention provides a kind of method for rapidly separating Geobacillus stearothermophilus spores and vegetative thalline, comprising the following steps:

[0042] In step one, the suspension containing spores and bacterial cells is collected in a sterile centrifuge container.

[0043]Step 2, put the centrifuge container into the centrifuge equipment for centrifugation, the centrifugation speed is selected as 7500-8500rpm, preferably 8500rpm in this embodiment, the centrifugation temperature is 4°C, and the centrifugation time is 10min, which can realize the suspension of vegetative bacteria and the separation of spores precipitation.

[0044] Step 3: Discard the upper liquid of the suspension after centrifugation, and add an appropriate amount of suspending medium to resuspend the sediment layer in the suspending medium. The suspending medium includes sterile purified water, suspending agent solution or density gradient separation liquid. In an example, a density g...

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Abstract

The invention discloses a method for rapidly separating Geobacillus stearothermophilus spores from nutrient thalli. The method comprises the following steps: S01, collecting a suspension containing spores and bacterial thalli into a sterile centrifugal container; S02, placing the centrifugal container into centrifugal equipment for centrifugation; S03, discarding upper layer liquid of centrifugedsuspension, and adding a proper amount of suspension medium to resuspend a precipitate layer in the suspension medium; S04, uniformly dispersing the precipitate layer in the suspension medium; and S05, after the suspension is uniformly dispersed, repeating the S02, the S03 and the S04 until the purity of the spores detected by a microscope meets the requirement. The method for rapidly separating the Geobacillus stearothermophilus spores from the nutrient thalli provided by the invention has the advantages of high spore separation efficiency, low time consumption and high separation purity.

Description

technical field [0001] The invention relates to a method for rapidly separating spores and vegetative cells of Geobacillus stearothermophilus, belonging to the technical field of microbial spore fermentation. Background technique [0002] In the field of disinfection research, microbial indicators should be used to evaluate the killing effect of various physical and chemical factors on pathogenic microorganisms. The effect of disinfection and sterilization equipment. The "Biological Indicators for Moist Heat Sterilization" GB18281.3-2015 issued by the Standardization Administration of China in 2015 was officially implemented on January 1, 2017. Officially changed Bacillus stearothermophilus to Geobacillus stearothermophilus. It belongs to thermophilic aerobic bacillus and is a Gram-positive bacterium. As a biological indicator standard bacterium, the spores produced are non-pathogenic, non-pyrogenic and non-toxic. The spore has definite resistance to certain sterilization...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/02C12R1/07
CPCC12N1/02C12N1/20
Inventor 王卫凌莉
Owner NANJING JUSHA DISPLAY TECH
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