Primer design method of multiple PCR for discriminating vaccine of tubercle branch bacillus

Abstract

This invention discloses a method for identifying multiple PCR primers of tuberculous mycobacteria strains, which comprises the steps of: (1) adding an abacerial genome sequence to 5' end of oligonucleotide primers P1 and P2 that can combine with the bacterium genome sequence to obtain specific long primers YB1-P1and YB2-P2 as the primer pair for the polymerase chain reaction in the first stage of multiplex amplification process; and (2) directly using the abacterial genome sequence as the peimer pair for the polymerase chain reaction in the first stage of multiplex amplification process.

Description

technical field [0001] The invention relates to a multiplex PCR primer design method for amplifying gene fragments of multiple bacteria at one time to identify Mycobacterium tuberculosis strains. Background technique [0002] Since the 1990s, with the rapid development of molecular biology in the medical field, researchers have also introduced molecular biology techniques into the scope of tuberculosis research. Polymerase chain reaction (PCR) is a molecular biology technique based on nucleic acid biochemistry, and it is one of the most valuable techniques in the field of biology. Since the diagnosis of tuberculosis was introduced in 1989, it has immediately become the focus of much attention in the field of tuberculosis bacteriological diagnosis. After years of scientific research and clinical testing, this method has been continuously improved, and its sensitive, rapid and specific characteristics have been verified in most reports. [0003] This project takes four clini...

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Problems solved by technology

The problems that these methods exist are: (1) strict PCR reaction conditions are needed, such as the concentration of various reaction components, the ratio between the reaction components, the temperature and time of PCR amplification, etc.

(2) Since a variety of different primers exist in the same reaction tube, the reaction between the primers is more obvious, such as the formation of primer dimers, etc., which will lead to the loss of a large amount of primers, which can easily lead to this multiple reaction. failure of the PCR reaction

(3) The selection of primers will be very strict, and it must be ensured that primer dimers will not form between them, which will greatly reduce the possibility of multiple PCR with different types of Mycobacterium tuberculosis

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