Method for preparing bacterial nucleic acid fingerprint characteristic spectrum library

Abstract

The invention discloses a method for preparing a bacterial nucleic acid finger print library, wherein the method comprises PCR amplification, SAP enzyme digestion, transcription, nucleic acid digestion through enzyme, purification, mass spectrometer detection and other steps. According to the present invention, based on the method, a nucleic acid finger print library of the common bacterial is established; and according to a mass spectrum characteristic spectrum library produced by experiments, classification and identification can be performed on bacterial to be detected, and results can be widely used for bacterial classification and identification, phylogenetic analysis, drug screening, import and export inspection and other fields.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for preparing bacterial nucleic acid fingerprints and a method for classifying and identifying bacteria using the method. Background technique [0002] The early bacteriological classification mainly adopted the traditional bacteriological classification method based on morphological and cultural characteristics and physiological and biochemical characteristics. However, because the surface characteristics of bacteria are often affected and limited by many human factors, especially the differences in people's subjective judgments, it is difficult to reflect the natural kinship of bacteria, which reflects the fact that bacteria are only based on morphological characteristics and physiological and biochemical characteristics. limitations of traditional taxonomies. [0003] Therefore, Colweel proposed a new bacterial taxonomy term "Polyphasic taxonomy", which refers to the metho...

AI Technical Summary

Problems solved by technology

Since this method uses conventional processing (treatment with absolute ethanol, formic acid and acetonitrile, supplemented by centrifugation, and finally aspirating the supernatant for detection), although it can characterize the characteristic map of the bacterium to a certain extent, it is still pending The test object contains proteins, lipids, lipopolysaccharides and lipooligosaccharides, DNA, polypeptides and other molecules that can be ionized. The amount of information in the map is too large, and the characteristic of the map is low due to the large number of molecules to be detected. It is only applicable to a specific bacterium and cannot be extended to other large numbers of bacterial detection.

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