A method for extracting bacterial metagenomic DNA from oral and pharyngeal swabs

Abstract

The present invention provides an oral throat swab bacterial metagenome DNA extraction method, which comprises five steps such as oral throat swab pretreatment, bacterial lysis, DNA separation and hybrid protein removing, adsorption of DNA through a silica gel membrane purification column and impurity removal, and elution of DNA so as to extract the oral throat swab bacterial genome DNA. According to the present invention, with the special sample treatment method, the yield can be substantially increased; the combination of ultrasonic wave crack cleavage and enzyme cleavage is used, and the appropriate wall breaking time and the mild conditions are used, such that the gram-negative bacteria, the gram-positive bacteria and other bacteria are subjected to appropriate wall breaking, and the breaking of the genomic DNA due to the excessive strong physical factors can not be produced; and the oral throat swab bacterial metagenome DNA can be extracted only in 1.5 h, and the bacterial metagenome yield is high.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for extracting bacterial metagenomic DNA from oral and throat swabs. Background technique [0002] Metagenome refers to the sum of all microbial genes in the habitat, which includes the sum of the genes of cultivated and uncultivated microorganisms. Humans live in a microbial world. As a complete human body, eukaryotic cells only account for 10% of the total number of cells, while 90% are prokaryotic cells. Therefore, Nobel Prize winner Lederberg proposed that the human body is symbiotic with eukaryotic cells and the body A "super organism" composed of prokaryotic cells. [0003] The human oral cavity and pharynx provide a good habitat for bacteria. Once the flora is out of balance, it will affect the health of the whole body. Scholars have proposed that bacterial communities may be the cause of some chronic diseases. The object of bacterial metagenomic resear...

AI Technical Summary

Problems solved by technology

[0004] The currently developed oropharyngeal swab extraction methods can only target one type of Gram-positive bacteria or Gram-negative bacteria, which lacks versatility, so the abundance of extracted metagenomic DNA is not good

In addition, although some methods can better extract the genomic DNA of bacteria from oropharyngeal swabs, they have the disadvantages of high cost, low yield, and cumbersome operations.

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