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Indoor micro-ecological environment livability detection method based on human conditioned pathogens

A detection method and technology for pathogenic bacteria are applied in the field of indoor micro-ecological environment livability detection based on human conditional pathogenic bacteria, which can solve the problems of long time and achieve the effect of rapid judgment.

Pending Publication Date: 2022-03-11
QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, the culture method is currently the "gold standard" for the detection and evaluation of the indoor micro-ecological environment. The bacteria that can be cultured and detected only account for less than 10% of the environmental flora. It can be seen that this method has limitations in detection and takes a long time. A method for detecting / evaluating the livability of the indoor micro-ecological environment by the content and diversity of the conditional pathogenic bacteria planted in the indoor flora environment

Method used

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  • Indoor micro-ecological environment livability detection method based on human conditioned pathogens
  • Indoor micro-ecological environment livability detection method based on human conditioned pathogens
  • Indoor micro-ecological environment livability detection method based on human conditioned pathogens

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: Collection of environmental flora samples

[0030] 1. Determine the designated sampling position in the process of collecting samples with swabs (for some clean environmental samples, ≥25cm 2 ), mark 16cm with a ruler / template 2 Area.

[0031] 2. Label all collection tubes.

[0032] 3. Put on rubber gloves, take the disposable swab out of the package, and be careful not to touch the surface of the swab with others.

[0033] 4. Immerse the swab head into the mixed solution of 0.01mol / L NaCl and 0.1% Tween20, and then press the swab into the tube to remove excess liquid.

[0034] 5. Smear the swab on the marked area horizontally and vertically (total 50 times, about 30-35s) for sampling.

[0035] 6. Cut off the swab head, put it into a marked, 2mL empty tube, and cap the tube tightly.

[0036] 7. Place the sample in an ice box, and then transfer it to a -80°C refrigerator for storage.

Embodiment 2

[0037] Example 2: Extract the DNA of the sample and perform high-throughput sequencing

[0038] 1. Pipette 350 μL of PBS solution and add it to the above 2 mL centrifuge tube containing the sample.

[0039] 2. Add 350 μL AL buffer, 40 μL lysozyme (10 mg / mL), 6 μL mutanolysin (25000 U), and 300 mg glass beads. Mix well, and incubate the sample at 37°C for 1h.

[0040] 3. Transfer the centrifuge tube to a tissue grinder (Qiagen) for 3 min at 26 Hz,

[0041] 4. Add 20 μL proteinase K (provided by QIAGEN kit) to the tube, and shake to mix.

[0042] 5. Incubate at 56°C for 3h.

[0043] 6. Transfer the supernatant to a new tube and discard the swab. with ddH 2 O wash the beads 2 times with 200 μL each.

[0044] 7. Add 1 / 2 volume of alcohol and mix well.

[0045] 8. Put the 2mL collection tube (provided in the kit) under the Dneasy centrifugal column, draw the mixture in step 7 to the spin column, centrifuge at 8000rpm / min for 1min, and discard the waste liquid and the collect...

Embodiment 3

[0050] Example 3: Using bioinformatics to analyze community structure

[0051] The off-machine data is based on the GreenGenes 16S rRNA full-length sequence database. Using software, the bacteria contained in the sample are classified, and the structure of the genus layer in the evolutionary classification is restored, and finally the relative abundance of the bacteria contained in the sample is obtained. further said to use QIIME software (Quantitative Insights Into MicrobialEcology, the coding sequence (barcode) of the original data is split, use Trimmomatic software to carry out the first quality control (parameter is set to Trimmomatic: SLIDINGWINDOW:30:25MINLEN:25), Use FLASH software to fuse the sequence data at both ends (parameter setting is FLASH: -M 200 -m 5 -x 0.1), and then use Fastx Toolkit software for secondary quality control (parameter setting is Fastx Toolkit: -Q 33 -q 25 -p 80). Finally, the QIIME software was used to remove the chimera (chimera), and finall...

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Abstract

The invention relates to the field of environmental ecological monitoring, in particular to an indoor micro-ecological environment livability detection method based on human conditioned pathogens. The detection method comprises an acquisition module, a detection module and a calculation module. The acquisition module is used for collecting a flora sample of a to-be-detected environment and extracting DNA of the flora sample; the detection module takes the sample DNA provided by the acquisition module as a raw material, and performs high-throughput sequencing to complete flora detection; and the calculation module analyzes and calculates the flora structure of the sample by using the sample sequencing data obtained by the detection module and a bioinformatics means to obtain an indoor micro-ecological environment index MiBE so as to judge whether a pathogenic risk exists in the current indoor environment or not. The method has the advantages of simple operation, no need of excessive pretreatment on samples, sensitive and rapid detection, and realization of the whole treatment and detection process within several hours, and the detection result has universality for yellow race and white race, and is an indispensable new quantification tool in the research of the field of indoor environments in public places.

Description

technical field [0001] The invention relates to the field of environmental ecological monitoring, in particular to a method for detecting the livability of an indoor micro-ecological environment based on human conditional pathogenic bacteria. Background technique [0002] In the daily life of modern society, people spend about two-thirds of the time in a relatively closed indoor environment, which often repeats: the three points and one line life of family-transportation-office also means that the external microorganisms that people come into contact with Mostly from the indoor environment. Among them, public places such as transportation hubs, shopping malls, farmers' markets, hospitals, and schools are crowded with people, and the air microorganisms are more complex. Microbes in the indoor air interact with the human body through various ways such as sedimentation into food, contact with human skin, or direct inhalation. Contact and interaction affect human microbes, whic...

Claims

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Application Information

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IPC IPC(8): C12Q1/6869C12Q1/06C12Q1/04
CPCC12Q1/6869C12Q2531/113C12Q2535/122C12Q2537/165
Inventor 徐健孙政朱鹏飞张丽丽代娅婕
Owner QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI