Method for detecting toxicity of organic pollutants in water body based on DNA damage repair defect living cell comet

A DNA damage and organic pollutant technology, applied in the field of environmental health genetic toxicology, to achieve the effects of improved sensitivity, simple operation and low cost

Active Publication Date: 2021-05-11
CHANGSHA UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] However, there is no technical method to detect the potential genotoxicity of organic pollutants in water by using living cells with DNA damage repair defects combined with single-cell gel electrophoresis experiments.

Method used

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  • Method for detecting toxicity of organic pollutants in water body based on DNA damage repair defect living cell comet
  • Method for detecting toxicity of organic pollutants in water body based on DNA damage repair defect living cell comet
  • Method for detecting toxicity of organic pollutants in water body based on DNA damage repair defect living cell comet

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Embodiment

[0037] A method for detecting the toxicity of organic pollutants in water bodies of the present invention comprises the steps of:

[0038] 1. Pretreatment of water samples

[0039] In June 2019, 20L of water samples were collected in the drinking water source section of the Xiangjiang River in Changsha City, Hunan Province (Wangcheng District) according to the standard method of the National Environmental Protection Agency. After standing still for 24 hours, the suspended particles were removed by filtration and enriched by XAD-2 resin (Sigma) adsorption column. The flow rate is 30-40mL / min, the water is removed with nitrogen, and then eluted with methanol, acetone and dichloromethane in sequence. The eluate is concentrated at 50°C, dried, and finally dissolved with dimethylsulfoxide (DMSO). Dilute to 2.0mL, and store at -18°C in the dark for later use.

[0040] 2. Cell Culture and Preparation

[0041] Construct gene homologous recombination targeting vector, transfect DT40...

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Abstract

The invention discloses a method for detecting toxicity of organic pollutants in a water body based on DNA damage repair defect living cell comet. The method comprises the following steps: collecting a water sample enrichment, treating DNA damage repair defect living cells (TDP1KO and TDP2KO) by the water sample enrichment while taking DNA damage repair normal living cells (WT) treated by the water sample enrichment as a contrast, combining with a single cell gel electrophoresis technology, analyzing and measuring DNA comet migration tail moment values of living cells and normal cells with DNA damage repair defects, and evaluating the toxicity of organic pollutants in a water body by using the DNA comet migration tail moment values. According to the method disclosed by the invention, the detection sensitivity of the genetic toxicity of the organic pollutants in the water body is improved, ultratrace organic pollutants in the water can be detected, the method has important significance on early warning of environmental pollution harming health, and a molecular mechanism of the biological toxicity of the organic pollutants in the water can be explored from the perspective of DNA damage repair.

Description

technical field [0001] The invention belongs to the field of genetic toxicology of environmental health, and in particular relates to a method for detecting the toxicity of organic pollutants in water bodies by using DNA damage repair defective living cells combined with single-cell electrophoresis experiment technology. Background technique [0002] With the increase of population and the rapid development of modern industry and agriculture, the problem of environmental pollution is becoming more and more serious. The results of epidemiological surveys at home and abroad show that there are organic pollutants in tap water and water source water. These organic substances can enter the human body through the water and food chain, causing human genetics Substance DNA damage affects the genetic stability of cells, causing miscarriage, stillbirth, teratogenicity, and the occurrence of certain genetic diseases and cancers. Therefore, it is very important to detect the toxicity of...

Claims

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Application Information

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Patent Type & AuthorityApplications(China)
IPC IPC(8): G01N21/64G01N27/447C12Q1/02
CPCG01N21/6428G01N27/447G01N33/5014G01N33/5052Y02A20/20
Inventor曾志红刘敏卓刘芳陈建荣陈珺阳宗欣
OwnerCHANGSHA UNIVERSITY