Sample pretreatment method for extracting microorganism total DNA in biological aerosol

Abstract

The invention relates to a sample pretreatment method for extracting microorganism total DNA in biological aerosol. The method includes the steps that a filter membrane is adopted to collect air microorganisms, 0.8% normal saline is used as a buffer solution, the microorganisms are fully separated under the oscillation action of a shaking table supplemented by glass beads, and membrane stripping liquid is filtered through a 25-mm filter membrane, so that the microorganisms are concentrated on the surface of a membrane with the area of 1.96 cm<2> and oscillated, walls of the microorganisms, with thick cell walls, on the surface of the membrane are broken preliminarily, and follow-up DNA extraction is facilitated. The sample pretreatment method is easy to operate, harmless to the human body and capable of effectively increasing the DNA extraction amount.

Description

technical field [0001] The invention relates to the research field of microorganisms in air aerosols, in particular to a sample pretreatment method for extracting total DNA of microorganisms in bioaerosols. Background technique [0002] Bioaerosols are aerosol particles with life (including microbial particles such as bacteria, fungi, and viruses), active particles (pollen, spores, etc.), and plasmids released into the air by living organisms. Because the concentration of bioaerosols in the air is low, the research on them first needs to collect the biological particles in the air so that the air microorganisms can be enriched. At present, there are three main collection methods: (1) using an impact sampler to make the bioaerosol hit the surface of the selective medium, and use it for subsequent analysis after the target air microorganisms grow; (2) using the sampler to collect the bioaerosol in the air Absorb into the buffer solution, filter the buffer solution with a filt...

AI Technical Summary

Problems solved by technology

However, the first method has a large filter membrane area, which leads to a large amount of DNA loss during the DNA extraction process; although the second method has a small loss during the extraction process, the buffer used in the stripping process, For example, 0.1% TritonX-100 buffer or phosphate buffer containing 0.03% Tween-20 contains organic substances harmful to human health, and the buffer configuration process is complicated and the steps are cumbersome