Method for extracting microbial DNA from sample

Abstract

A method for extracting microorganism DNA from a sample mainly comprises the steps of sample liquefaction and pathogenic microorganism nucleic acid extraction, liquefaction comprises the step of adding a liquefaction reagent and Benzonase nuclease, and simultaneous addition of the Benzonase nuclease not only can doubly liquefy the sample, but also can remove part of host nucleic acid and shorten metagenome extraction time, and the extraction time of the sample is shortened. According to the method, the viscosity of the sample can be more efficiently and fully reduced, nucleic acid pollution of damaged host cells in the sample is removed, the time cost is greatly saved, and rapid detection of metagenomes is facilitated.

Description

technical field [0001] The invention relates to the field of microbial molecular detection, in particular to a method for extracting microbial DNA from samples, the main samples including sputum and abscess fluid. Background technique [0002] Infections caused by respiratory diseases mainly involve sputum samples, and the common infectious pathogenic microorganisms include Acinetobacter baumannii, Streptococcus pneumoniae, Mycobacterium tuberculosis, Moraxella catarrhalis, Pseudomonas aeruginosa, Aspergillus fumigatus, black Aspergillus herpes simplex virus, etc. These pathogenic microorganisms are mostly infectious and need to be strictly operated in a biological safety cabinet, and general laboratories do not have operating equipment. Common sputum samples are relatively viscous and need to be liquefied to destroy the cohesin in the sputum sample. At present, the commonly used liquefaction reagents are sodium hydroxide, dithiothreitol, N-acetic acid-L-cysteine In 2009, Z...

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Problems solved by technology

However, these methods can only detect one or a limited number of pathogens at a time. In addition, the culture still has the disadvantage of low positive rate, and most of the abscess fluid collected in clinical samples is relatively viscous, and the sample is turbid as a whole, and precipitated after high-speed centrifugation The separation is not obvious, and it is not easy to remove the supernatant when discarding it. The purity of the nucleic acid product extracted from the obtained precipitate is not high and most of it is host nucleic acid

The emerging metagenomic detection technology (mNGS) can sequence the genome of microbial populations in a specific environment, avoiding the defects that most microorganisms cannot be cultured in traditional methods, but for the problem of a high proportion of human DNA, it is the detection of pathogenic microorganisms bring difficulty

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