Method for detecting biological toxicity of atmospheric particulates

A technology for real-time detection of atmospheric particulate matter, applied in measurement devices, material excitation analysis, fluorescence/phosphorescence, etc., to achieve the effect of high sensitivity and low detection limit

Inactive Publication Date: 2017-01-04
PEKING UNIV
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  • Claims
  • Application Information

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  • Method for detecting biological toxicity of atmospheric particulates
  • Method for detecting biological toxicity of atmospheric particulates
  • Method for detecting biological toxicity of atmospheric particulates

Examples

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Embodiment 1

[0027] Embodiment 1. Detection of biological toxicity of atmospheric particulate matter

[0028] (1) Use Teflon sampling membrane and four-channel atmospheric particle sampler to collect PM2.5 particulate matter in Beijing atmosphere, and the sampling time is 24 hours. Weigh the mass of atmospheric particulate matter on the sampling membrane and dissolve it in ultrapure aqueous solution to make a particulate matter solution with a concentration of 0.04mg / mL;

[0029] (2) Add the above-mentioned particle solution solution into a 96-well plate, and add the yeast liquid that utilizes GFP-labeled control SSA1 and HSP60 protein expression gene fragments;

[0030] (3) Put the 96-well plate into the microplate reader, and use the microplate reader to dynamically monitor the change of the fluorescence value of each experimental group in the 96-well plate. The microplate reader reads a fluorescence value every 1 minute, and the reading time is 2 hours, the reading wavelength is 480-52...

Embodiment 2

[0037] Example 2. Detection of biological toxicity of endotoxin

[0038] (1) Dilute the purchased endotoxin sample with a concentration of 50ng / mL to 0.5ng / mL for later use;

[0039](2) adding the above-mentioned endotoxin solution into a 96-well plate, and respectively adding yeast liquids for controlling SSA1 and HSP60 protein expression gene fragments labeled with GFP;

[0040] (3) Put the 96-well plate into the microplate reader, and use the microplate reader to dynamically monitor the change of the fluorescence value of each experimental group in the 96-well plate. The microplate reader reads a fluorescence value every 1 minute, and the reading time is 2 hours, the reading wavelength is 480-520nm;

[0041] (4) Analyze the fluorescence value recorded by the microplate reader, and the fluorescence value detection result after 2 hours is as follows: image 3 and Figure 4 shown.

[0042] Experimental results:

[0043] 1. From image 3 It can be seen that compared with ...

Embodiment 3

[0047] Embodiment 3. Compare the biotoxicity of different urban atmospheric particles

[0048] 1. Use the four-channel high-flow atmospheric particle sampler and TEFLON sampling membrane to collect PM2.5 samples from different cities such as Beijing, Guangzhou, Lanzhou, and Haikou.

[0049] 2. Make the collected samples into a particle solution with a concentration greater than 0.02mg / mL for later use.

[0050] 3. Add the above-mentioned particle solution into a 96-well plate, and add the yeast liquid that uses GFP-labeled gene fragments to control the expression of SSA1 and HSP60 proteins.

[0051] 4. Put the 96-well plate into the microplate reader, and use the microplate reader to dynamically monitor the change of the fluorescence value of each experimental group in the 96-well plate. The microplate reader reads a fluorescence value every 1 minute, and the reading time is 2 hours , the reading wavelength is 480-520nm.

[0052] 5. Analyze the fluorescence value of the part...

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Abstract

The invention discloses a method for detecting the biological toxicity of atmospheric particulates. The method comprises the steps that two proteins SSA1 and HSP60 havingspecific sensitive response to oxidative stress of saccharomycetes are screened, a gene fluorescence probe is utilized to mark and control genes of the specific proteins of a living saccharomycetes body to make the living saccharomycetes body serve as an atmospheric particulate indicator microbe, and the biological toxicity of the atmospheric particulates is inversed by detecting the expression concentrations of different fluorescent proteins having specific wavelength in real time. The toxicity of atmospheric particulates is detected at the level of the single living saccharomycetes body, the method has the advantages of being high in sensitivity, low detection limit and the like, can be applied to thehuman health effectassessment of the atmospheric particulates and biological ingredients and measurement and real-time monitoring of air quality.

Description

technical field [0001] The present invention relates to an analysis technique for detecting the biological toxicity of atmospheric particles in real time. Specifically, a gene fluorescent probe is used to mark a gene controlling a specific protein related to oxidative stress in living yeast, so that living yeast can be used as an indicator microorganism for the toxicity of atmospheric particles. Real-time detection of different fluorescent protein expression concentrations at specific wavelengths to retrieve the biological toxicity of atmospheric particles can be applied to the assessment method of the health effects of atmospheric particles and biological components on the human body. Background technique [0002] In recent years, severe smog has occurred frequently in many places in China, and a lot of work has been devoted to studying the mechanism of its formation, among which coal combustion and vehicle exhaust are listed as important contributors. Atmospheric particula...

Claims

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Application Information

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IPC IPC(8): G01N21/64
CPCG01N21/6428G01N2021/6439
Inventor 要茂盛魏恺
Owner PEKING UNIV
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