Method for rapidly detecting and analyzing multi-copy number of MIRU-VNTR genes of mycobacterium tuberculosis

Abstract

The invention provides a method for rapidly detecting and analyzing the multi-copy number of MIRU-VNTR genes of mycobacterium tuberculosis. By carrying out PCR amplification and sequencing on a targetgene and calculating the copy number, a sequenced VNTR gene sequence is analyzed by virtue of EditSeq software, and the occurrence number of a repeated fragment is recorded. Compared with a traditional calculation manner, the method has the advantages that the operation time for calculating the repetition number of the VNTR of the mycobacterium tuberculosis is short, a result is relatively precise, and the operability is relatively strong. Used instruments are PCR and gel electrophoresis instruments and EditSeq open software of a computer analysis system, and operation techniques are relatively mature. By analyzing the copy number of the VNTR gene sequence, the epidemiology of the mycobacterium tuberculosis can be rapidly diagnosed. By rapidly and precisely analyzing the copy number of the VNTR, the epidemic condition of the mycobacterium tuberculosis is researched, thereby being beneficial to rapid clinical preliminary screening and community screening of tuberculosis.

Description

technical field [0001] The invention belongs to the field of rapid calculation of microbial gene copy numbers, and in particular relates to a rapid detection and analysis method for multiple copy numbers of Mycobacterium tuberculosis MIRU-VNTR genes. Background technique [0002] Tuberculosis (TB) is a chronic infectious disease caused by Mycobacterium tuberculosis (MTB) infection, and the main clinical manifestation is pulmonary tuberculosis. Most infected persons will finally show clinical symptoms after a long incubation period due to the persistent immune response of the body to MTB antigen stimulation. [0003] Early and rapid diagnosis of tuberculosis has always been the key to the prevention and treatment of tuberculosis. Traditional laboratory tuberculosis detection methods have their own defects. With the development of nucleic acid amplification technology in recent years, molecular biology detection is being more and more widely used clinically. [0004] Molecul...

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Problems solved by technology

However, this method is cumbersome. If the band amplification is not obvious and not bright enough during gel electrophoresis, the results cannot be displayed, and the experimental results are easily affected by subjective factors.

2. Formula calculation method, the commonly used formula at present: repeat sequence copy number = (PCR product fragment length - flanking sequence length) / repeat sequence length, but this method is relatively cumbersome and the calculation results are not accurate. Currently, Mycobacterium tuberculosis is reported in the literature There are few articles on flanking sequences, and they are not authoritative, and the values ​​​​are not accurate

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