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Genetic engineering biological indicator

A biological indicator and genetic engineering technology, applied in the field of disinfection and sterilization biological indicators, can solve the problem of inconsistency between enzyme heat resistance and spore heat resistance.

Active Publication Date: 2016-08-31
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

[0033] The genetic engineering biological indicator is composed of the following parts: Bacteria sheet, which is evenly distributed with genetic engineering indicator microorganisms.

[0034] 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.

[0035] 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.

[0036] 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

[0038]The genetic engineering biological indicator is composed of the following parts: Bacteria sheet, which is evenly distributed with genetic engineering indicator microorganisms.

[0039] 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.

[0040] 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.

[0041] 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

[0043] The genetic engineering biological indicator is composed of the following parts: bacteria sheet, which is evenly distributed with genetic engineering indicator microorganisms.

[0044] 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.

[0045] 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.

[0046] 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 downs...

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PUM

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Abstract

The invention discloses a genetic engineering biological indicator used for biological monitoring of one or more sterilization effects. According to the genetic engineering biological indicator, foreign genes, including promoters, reporter genes and selectable maker genes, are guided into indicator microorganisms by means of genetic engineering. The selectable maker genes are positioned on the upstream of the inducible promoters, and the reporter genes are positioned on the downstream of the inducible promoters. In the absence of inducers, the selectable maker genes are expressed, the reporter genes are not expressed, and accordingly, successfully-transfected strains are screened. After exposure by sterilization factors, the indicator microorganisms contact with the inducers, and the reporter genes are expressed. Autofluorescence of the genetic engineering biological indicator can be achieved without specific enzyme substrates, and the genetic engineering biological indicator has the advantages of high signal-to-noise ratio, strong fluorescence signal, easiness in detection, high sensitivity and the like.

Description

Technical field [0001] The present invention involves a disinfected sterilizer biosensory agent, especially for a genetic engineering indicator for one or more sterilization effect and for biological monitoring. Background technique [0002] In the healthcare industry, the health quarantine industry, pharmaceutical companies and other industries involving sterile technology, it is often necessary to evaluate and verify the sterilization effects of various sterile production processes, sterile products, sterile packaging materials, and test equipment.The sterilization effect.Every year, there are reports that cause infection due to incomplete sterilization.Biological testing is a standardized strain, and its required resistance to evaluate whether the sterilization load meets the requirements of sterile protection. [0003] The currently widely used biosensors at home and abroad are universal bioscents and fast biosensors.General -purpose biosensors are based on the recovery of sp...

Claims

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Application Information

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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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