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Novel genetic engineering biological indicator

A biological indicator and genetic engineering technology, applied in the field of new genetic engineering biological indicators, can solve the problems of low efficiency and low accuracy of exogenous gene preparation, and achieve the effects of easy operation, simplified preparation process and high preparation success rate

Inactive Publication Date: 2017-04-26
NANJING JUSHA DISPLAY TECH +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the problems of low preparation efficiency and low accuracy of exogenous genes in the prior art, the present invention provides a novel genetic engineering biological indicator

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] The genetically engineered biological indicator includes a filter paper and an ampoule sealed in the package. Genetically engineered Bacillus stearothermophilus is evenly distributed on the filter paper. The interior of the ampoule is sealed with 2ml of soy peptone liquid medium. The packaging is used to ensure that the filter paper is in place during storage. In the sterile environment, the sterilizing factor is in contact with the filter paper during the sterilization process, and the soy peptone liquid medium in the ampoule is separated from the filter paper. After the sterilization is completed, the soy peptone liquid medium is in contact with the filter paper.

[0029] The foreign genes introduced by the genetic engineering Bacillus stearothermophilus include alkaline phosphatase promoter, ampicillin resistance gene and enhanced green fluorescent protein EGFP reporter gene. The foreign genes are alkaline phosphatase promoter, ampicillin resistance gene and enhanced gre...

Embodiment 2

[0031] The genetic engineering biological indicator includes a stainless steel sheet and an ampoule sealed in the package. The genetic engineering Bacillus subtilis is evenly distributed on the stainless steel sheet. The inside of the ampoule is sealed with 2ml nutrient broth medium. The packaging is used to ensure that the stainless steel sheet is in In the sterile environment, the sterilizing factor is in contact with the stainless steel sheet during the sterilization process, and the nutrient broth medium in the ampoule is separated from the stainless steel sheet. After the sterilization is completed, the nutrient broth medium contacts the stainless steel sheet.

[0032] The foreign genes introduced by the genetic engineering Bacillus subtilis include alpha-L-arabinofuranosase promoter, LacZ blue-white spot screening gene and blue fluorescent protein BFP reporter gene. The foreign genes from upstream to downstream are the alpha-L-arabinofuranosidase promoter, the LacZ blue-whit...

Embodiment 3

[0034] Genetically engineered biological indicators include plastic tablets and ampoules sealed in the packaging. The plastic tablets are evenly distributed with genetically engineered Bacillus subtilis black variant spores. The ampoules are sealed with 0.5% by mass glucose broth medium. The packaging is used to ensure The plastic sheet is stored in a sterile environment and the sterilization factor is in contact with the plastic sheet during the sterilization process, and the 0.5% glucose broth medium in the ampoule is separated from the plastic sheet, and the 0.5% glucose broth is cultured after the sterilization is completed. The base is in contact with the plastic sheet.

[0035] The exogenous genes introduced into the genetically engineered Bacillus subtilis black variant spores include leucine aminopeptidase promoter, neomycin resistance gene and yellow fluorescent protein YFP reporter gene. The foreign genes from upstream to downstream are leucine aminopeptidase promoter, ...

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Abstract

The invention discloses a novel genetic engineering biological indicator. A foreign gene comprising a constitutive promoter, a reporter gene and a selectable maker gene is imported into an indicator microorganism through a gene engineering technique means, both the reporter gene and the selectable maker gene are located at the downstream of the constitutive promoter and are regulated and controlled by the upstream constitutive promoter, the selectable maker gene is used for screening a strain being successfully transfected, the reporter gene is used for indicating the growth of the microorganism after sterilization and exposure, and when the strain grows, the constitutive promoter is activated to promote the expression of the reporter gene and the selectable maker gene; according to the novel genetic engineering biological indicator provided by the invention, the constitutive promoter activated in the earlier stage of spore recovery growth is used for starting the expression of the foreign gene, remaining living spores can be quickly detected at the earlier stage of spore recovery growth, and the indicator is suitable for detection on the organism with one or more sterilization efficacies. The indicator provided by the invention has no need of any inductors or specific enzyme substrates, has a simple foreign gene system, is easy to operate and high in sensitivity and has a good industrial application prospect.

Description

Technical field [0001] The invention belongs to the field of disinfection and sterilization biological indicators, and particularly relates to a novel genetic engineering biological indicator. Background technique [0002] Biological testing is to evaluate whether the sterilization load meets the requirements of sterility assurance level through standardized strains whose resistance meets the requirements, and biological indicators used to detect or determine the effect of sterilization treatment came into being. [0003] Existing biological indicators are mainly divided into general-purpose biological indicators and rapid-type biological indicators. The general-purpose biological indicators are based on the acidic metabolites produced by the metabolism of spores when the spores resume growth and cause the pH of the medium to change. By adding pH indicators such as Bromocresol purple can be interpreted according to its color change. The microbial culture time is 24-48h. The result ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/22C12Q1/04
Inventor 王卫周平乐
Owner NANJING JUSHA DISPLAY TECH
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