Method for effectively enriching air medium microorganisms on basis of magnetic bead method

An air medium and microorganism technology, applied in the biological field, can solve problems such as difficult tracking, difficult quantification, and difficult enrichment, and achieve the effect of improving detection limit, high recovery rate and low cost.

Inactive Publication Date: 2015-10-07
HARBIN INST OF TECH SHENZHEN GRADUATE SCHOOL
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  • Application Information

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Problems solved by technology

[0004] Aiming at the technical problem that air medium microorganisms are difficult to enrich, the present invention provides a method for efficiently enriching air medium microorganisms based on the magnetic bead method, which can obtain high enrichment, high concentration, and easy-to-detect air medium under the premise of low cost Microorganisms, which effectively solve the problems of difficult tracking, enrichment and quantification in the process of microbial monitoring and analysis in air media

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  • Method for effectively enriching air medium microorganisms on basis of magnetic bead method
  • Method for effectively enriching air medium microorganisms on basis of magnetic bead method
  • Method for effectively enriching air medium microorganisms on basis of magnetic bead method

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Embodiment 1

[0043] Such as figure 1 As shown, a method for efficiently enriching air medium microorganisms based on the magnetic bead method specifically includes the following steps:

[0044] A: air medium microbial collection

[0045] (1) Sampling equipment: a large flow TSP (PM10) sampler produced by Qingdao Laoshan Applied Technology Research Institute, collect air samples and collect them on a 230mm×180mm glass fiber membrane filter;

[0046] (2) Sampling time: 10h, the maximum collection flow is 1.05m 3 / h;

[0047] (3) Sampling location: chicken farm.

[0048] B: Using the magnetic bead method to enrich microorganisms in the air medium

[0049] (1) Cut up the sampling filter membrane and place it in a beaker, take 1 / 4 of the filter membrane and use tweezers to carefully tear off the front adsorption layer of the filter membrane and place them in three 50mL centrifuge tubes;

[0050] (2) Add citric acid-disodium hydrogen phosphate buffer solution to the three centrifuge tubes, ...

Embodiment 2

[0074] A method for efficiently enriching air medium microorganisms based on a magnetic bead method, specifically comprising the following steps:

[0075] 1. Air medium microbial collection

[0076] (1) Sampling equipment: the large flow TSP (PM10) sampler produced by Qingdao Laoshan Applied Technology Research Institute, collects air samples and collects them on a 230mm×180mm glass fiber membrane filter membrane; the schematic diagram of the quantitative verification smoke chamber device is as follows image 3 shown;

[0077] (2) Sampling time: 10h, the maximum collection flow is 1.05m 3 / h;

[0078] (3) Sampling location: 10m 3 Controllable quantitative verification smoke box, set the bacterial concentration to 10 6 cell / m 3 ;

[0079] (4) Enrichment method: the sampling membrane was processed by the magnetic bead method and the buffer dispersion method respectively.

[0080] 2. Enrichment of microorganisms in air medium by magnetic bead method

[0081] (1) Cut up th...

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Abstract

The invention provides a method for effectively enriching air medium microorganisms on the basis of a magnetic bead method. The method comprises the following steps: placing a filter membrane in a to-be-detected environment, performing the sampling in an adsorption manner, and placing the filter membrane after the sampling into an extraction container; adding an acid buffer solution with pH value of 2.0 to 2.5, then oscillating for 5 to 10 minutes, and then carrying out the ultrasonic dispersion for 10 to 20 minutes; adding ferroferric oxide magnetic beads into the extraction container, and continuously ultrasonically dispersing for 10 to 30 seconds; taking another separation container, charging the acid buffer solution into the separation container, pouring the ultrasonically-dispersed solution into the separation container, oscillating, uniformly mixing, standing for 1 to 10 minutes, and then separating magnetic beads to obtain separated magnetic beads. By adopting the technical solution, the air medium microorganisms with high enrichment amount, high concentration and convenience in detection can be obtained on the premise of low cost, so that the problem of difficulty in tracking, enriching and quantifying in the monitoring and analysis process of air medium microorganisms can be effectively solved.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for efficiently enriching air medium microorganisms based on a magnetic bead method. Background technique [0002] Since the implementation of the "Aerosol Infection" program in the United States in 1943, microbial aerosols have been widely concerned at home and abroad. Bacteria in microbial aerosols include non-pathogenic bacteria, pathogenic bacteria and conditional pathogenic bacteria, which can cause human infection through airborne transmission to a certain extent, and even a very small amount of pathogenic bacteria can directly cause respiratory tract infection. Therefore, strengthening research on aerosol microbes closely related to humans is crucial to controlling human health risks. [0003] At present, the common detection methods of airborne microorganisms can be divided into two categories: culture method and non-culture method. The traditional cult...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/00C12N15/10
Inventor 邵明非高新磊李继张凯张彬声
Owner HARBIN INST OF TECH SHENZHEN GRADUATE SCHOOL
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