Method for identifying mycobacterium

Abstract

The invention discloses a method for identifying mycobacterium. The method comprises the following steps: acquiring a plurality of standard sequences of a plurality of mycobacteria so as to generate a mycobacterium sequence database; by comparing the standard sequences, obtaining a conservation section mutually owned by the standard sequences; preparing a PCR primer according to the conservation section; using the PCR primer to perform the PCR amplification of a DNA solution of the testing mycobacterium so as to obtain an amplification product of the testing mycobacterium; carrying out the sequencing reaction of the amplification product to obtain the sequencing product; carrying out the sequencing analysis of the sequencing product to obtain a sequence of the testing mycobacterium; and by comparing the testing sequence with the standard sequence in the mycobacterium sequence database, determining the type of the testing mycobacterium. The PCP primer is designed according to the conservation section of the standard sequences of various mycobacteria, only a pair of primers is needed for all PCRs and sequencing, thereby greatly reducing the quantity of oligonucleotide needed for experimental population.

Description

technical field [0001] The invention relates to clinical molecular diagnosis technology in the field of biotechnology, in particular to a method for identifying mycobacteria using PCR sequencing. Background technique [0002] The number of mycobacteria currently stands at about 110 species, and it is the cause of many diseases collectively known as mycobacterial diseases. Therefore, the classification and identification of mycobacteria has attracted the attention of many scientific researchers. . In the prior art, the identification techniques of mycobacteria are divided into three categories. [0003] One is to use conventional biochemical indicators to identify mycobacteria, such as described in patent CN1298023. This method is also a commonly used method in the current laboratory department. This method takes a long time to identify (needs 1-2 weeks), and the results of the identification are affected by human observation, so the accuracy is not very high. [0004] Th...

AI Technical Summary

Problems solved by technology

This method is also commonly used in clinical laboratories. This method takes a long time to identify (1-2 weeks), and the results of the identification are affected by human observation, so the accuracy is not very high.

[0004] The second type uses the antigen-antibody method for serological diagnosis. For example, the patent CN1388378 uses this method. The number of mycobacteria that can be detected by this method is also relatively small, and each bacterium requires an antibody for detection, so it cannot identify clinically relevant All mycobacteria

Therefore, it is not conducive to the application of clinical standardization

Moreover, the number of mycobacteria detected by chip hybridization method is limited

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