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Biological membrane DGT device and method for simultaneously and quantitatively monitoring effective state content and cell genetic toxicity of target

A genotoxic and biofilm technology, applied in the field of environmental science, can solve the problems of large differences in film-making materials, the inability of the diffusion membrane to meet the diffusion coefficient of heavy metals, and the loss of measuring the effective state content of heavy metals, and achieve the effect of simple and convenient assembly steps.

Inactive Publication Date: 2019-04-05
南京维申环保科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although the method of this application proposes the concept of combining biological probes with membranes, the adsorption membrane and diffusion membrane in the DGT device in the prior art are quite different from the membrane materials in this application. None of the materials can meet the survival requirements of microorganisms; if the adsorption membrane in the DGT device is modified with the above-mentioned membrane materials to meet the growth requirements of microorganisms, the DGT device will lose the function of measuring the effective content of heavy metals; if the above-mentioned membrane materials are used for Modified to meet the needs of microbial growth, the diffusion coefficient of the diffusion film cannot meet the requirements of the diffusion coefficient when monitoring the effective content of heavy metals, so it is difficult to combine microbial film sensors with DGT technology

Method used

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  • Biological membrane DGT device and method for simultaneously and quantitatively monitoring effective state content and cell genetic toxicity of target
  • Biological membrane DGT device and method for simultaneously and quantitatively monitoring effective state content and cell genetic toxicity of target
  • Biological membrane DGT device and method for simultaneously and quantitatively monitoring effective state content and cell genetic toxicity of target

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Experimental program
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Effect test

Embodiment 1

[0045] This embodiment is the preparation of the biosensor diffusion film, the assembly and sampling of the biofilm DGT device.

[0046] The preparation method of the biosensor diffusion film comprises the following steps:

[0047] 1) First prepare the suspension containing the biosensor ADPWH_recA: a) The luxCDABE gene is fused into the chromosome of the Acinetobacter baylyi ADP1 host cell to construct; b) The constructed biosensor needs to be cultured with 10 μg / mL kanamycin Culture medium (Luria-Bertani, LB) at constant temperature (30°C) for 16 hours; c) The biosensor in each 1 mL of the above medium was recovered by centrifugation (3000 rpm, 10 minutes), and the recovered biosensor ADPWH_recA was redissolved in 10mL of LB medium for later use;

[0048] 2) Add 1.1 g of agarose powder purchased from Sigma-Aldrich into 100 mL of deionized water, and mix to obtain an agarose solution;

[0049] 3) Put the agarose solution in step 2) into an autoclave, and heat at 121° C. for...

Embodiment 2

[0056] The present embodiment is basically the same as Example 1, except that the quality of the agarose powder in step 2) is 1.25 g; the mixing ratio in step 4) is 10:1 (v / v); cooling at room temperature in step 5) The time is 1.5 hours.

Embodiment 3

[0058] This embodiment is basically the same as Example 1, except that: this embodiment is basically the same as Example 1, except that the quality of the agarose powder in step 2) is 1.2g; the mixing ratio in step 4) is 9 : 1 (v / v); the cooling time at room temperature in step 5) was 1.2 hours.

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Abstract

The invention discloses a biological membrane DGT device and a method for simultaneously and quantitatively monitoring the effective state content and the cell genetic toxicity of a target, and belongs to the field of environment science. A diffusion membrane in the biological membrane DGT device is an agarose hydrogel tablet in which a biosensor is attached. The biological membrane DGT device isadopted for carrying out target collection; the effective state content and the cell genetics / genetic toxicity of the target are determined after pretreatment; the cell genetic toxicity is determinedby determining a fluorescence response value of the target in the diffusion membrane; and the cell genetic toxicity is analyzed by using a FLUOstar Omega microplate reader, so that the operation is simple and convenient, and the consumed time is short. The combination of the biosensor and a DGT technology is realized for the first time, so that the effective state content and the cell genetic toxicity of the target can be simultaneously and quantitatively monitored.

Description

technical field [0001] The invention belongs to the field of environmental science, and more specifically relates to a biofilm DGT device and a method for quantitatively monitoring the effective state content of a target substance and the genotoxicity of cells at the same time. Background technique [0002] In recent years, heavy metal pollution in the environment has aroused widespread concern in society. Excessive heavy metals can not only affect the growth of microorganisms and plants in the environment, but also cause irreversible damage to human health through the food chain or other channels (such as drinking water, breathing, etc.). Different from organic pollutants, the non-degradable characteristics of heavy metals cause their impact on the surrounding environment to become a long-term effect, so human intervention-remediation is necessary to gradually eliminate their threat to the environment. [0003] The current monitoring methods for heavy metals mainly include...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/62G01N21/64G01N21/31
CPCG01N21/31G01N21/6486G01N27/62
Inventor 吴双成皓
Owner 南京维申环保科技有限公司
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