genetically engineered biological indicator

A biological indicator and genetic engineering technology, applied in the field of disinfection and sterilization biological indicators, can solve the problems of inconsistent heat resistance of enzymes and spores

Active Publication Date: 2018-01-12
NANJING JUSHA DISPLAY TECH +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If the repressor gene is reduced, the gene will also be expressed under the condition of non-induction, and there will also be a phenomenon that the thermotolerance of the enzyme is inconsistent with the thermotolerance of the spore

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] The genetic engineering biological indicator is composed of the following parts: Bacteria sheet, which is evenly distributed with genetic engineering indicator microorganisms.

[0033] The inner packaging ensures that the bacteria slices are in a sterile environment during storage, and at the same time, the effective sterilizing factor can fully contact the bacteria slices during the sterilization process.

[0034] The ampoule bottle is sealed with 2ml soybean peptone liquid medium and 100mM IPTG inducer. The soybean peptone liquid medium and IPTG inducer are separated from the bacteria sheet during the sterilization process and allowed to contact with the bacteria sheet after the sterilization.

[0035] The genetic engineering indicator microorganism contains: 1*10 6 cfu~5*10 6 cfu Bacillus stearothermophilus spores, the exogenous gene is transfected into Bacillus stearothermophilus by pUB110 plasmid. The exogenous genes include the ampicillin resistance gene, locate...

Embodiment 2

[0037] The genetic engineering biological indicator is composed of the following parts: Bacteria sheet, which is evenly distributed with genetic engineering indicator microorganisms.

[0038] The inner packaging ensures that the bacteria slices are in a sterile environment during storage, and at the same time, the effective sterilizing factor can fully contact the bacteria slices during the sterilization process.

[0039] The ampoule bottle is sealed with 2ml nutrient broth medium and 100mM indole acrylic acid inducer inside, the nutrient broth medium and indole acrylic acid inducer are separated from the bacteria sheet during the sterilization process, allowing the same bacteria sheet contact.

[0040] The genetic engineering indicator microorganism contains: 1*10 6 cfu~5*10 6 cfu Bacillus subtilis spores, the exogenous gene is transfected into Bacillus subtilis by pSBPTQ plasmid. The exogenous genes include the LacZ blue-white screening gene, which is located downstream of ...

Embodiment 3

[0042] The genetic engineering biological indicator is composed of the following parts: bacterial flakes, which are evenly distributed with genetic engineering indicator microorganisms.

[0043] The inner packaging ensures that the bacteria slices are in a sterile environment during storage, and at the same time, the effective sterilizing factor can fully contact the bacteria slices during the sterilization process.

[0044] The ampoule bottle is sealed with 0.5% glucose broth medium and arabinose inducer in its inner seal. The 0.5% glucose broth medium and arabinose inducer are separated from the bacteria sheet during the sterilization process and allowed to contact with bacteria.

[0045] The genetic engineering indicator microorganism contains: 1*10 6 cfu~5*10 6 cfu Bacillus subtilis black mutated spores, the exogenous gene is transfected into the subtilis black mutated spores by pIH41 plasmid. The exogenous genes include the neomycin resistance gene, which is located do...

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PUM

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Abstract

The invention discloses a genetically engineered biological indicator, which is used for biological monitoring of one or more sterilization effects. It introduces an exogenous gene into the indicator microorganism by means of genetic engineering technology, and the exogenous gene includes a promoter. genes and selectable marker genes. The selectable marker gene is located upstream of the inducible promoter, and the reporter gene is located downstream of the inducible promoter. In the absence of inducers, the selectable marker gene is expressed and the reporter gene is not expressed, which is used to screen for successful transfection strains. After exposure to the sterilizing agent, the indicator microorganism is contacted with the inducer, and the reporter gene is expressed. The invention does not need a specific enzyme substrate, can autofluoresce, and has multiple advantages such as high sex-to-noise ratio, strong fluorescence signal, easy detection, and high sensitivity.

Description

technical field [0001] The invention relates to the field of disinfection and sterilization biological indicators, in particular to a genetically engineered biological indicator for one or more sterilization effects and for biological monitoring. Background technique [0002] In the medical care industry, health quarantine industry, pharmaceutical companies and other industries involving aseptic technology, it is often necessary to evaluate and verify the sterilization effects of various aseptic production processes, aseptic products, aseptic packaging materials, etc., and test equipment sterilizing efficacy. There are reports of infection incidents caused by incomplete sterilization every year around the world. Biological testing is to assess whether the sterilization load meets the requirements of the sterility assurance level through standardized strains and their resistance to the requirements. [0003] At present, the biological indicators widely used at home and abro...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12M1/34C12R1/125C12R1/07C12R1/145
CPCC12Q1/04C12Q2304/60C12M1/34C12N1/205C12R2001/07C12R2001/125C12R2001/145
Inventor 王卫周平乐
Owner NANJING JUSHA DISPLAY TECH
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