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Method for identifying mycobacterium piperculosis and nontuberculous mycobacterium

A technology of mycobacterium tuberculosis and mycobacteria, which is applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of inability to distinguish interspecies sequence NTM, inability to NTM typing, etc.

Pending Publication Date: 2021-10-15
新起点生物科技苏州有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The main problem of the present invention is that the existing identification technology can only identify a part of common pathogenic NTMs, and cannot distinguish a small amount of NTMs due to the high homology of interspecies sequences, and the existing identification technology cannot carry out more detailed classification of NTMs. Type and other issues

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  • Method for identifying mycobacterium piperculosis and nontuberculous mycobacterium
  • Method for identifying mycobacterium piperculosis and nontuberculous mycobacterium
  • Method for identifying mycobacterium piperculosis and nontuberculous mycobacterium

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Embodiment 1

[0060] The embodiment of the present invention provides a kind of identification method of mycobacterium tuberculosis and non-tuberculosis mycobacterium, please refer to Figure 1 to Figure 15 , including the following steps:

[0061] What needs to be explained first is that the target detection strains selected in this embodiment are as follows: Figure 5 and Image 6 As shown, the designed sequence information is as Figure 7 shown.

[0062] Homologous genes / sequences 16srRNA, 16s-23s rRNA interspace (ITS), mku, mce3B were selected.

[0063] Obtain the target detection strain, analyze the gene sequence information of the target detection strain, and determine the design coordinates.

[0064] For the detection of two different SNPs from A / G, design PCR primers and extension primers. Both ends of the upstream and downstream primers of the PCR primers need to add common sequences, namely ACGTTGGATG. The main purpose is to make the PCR primers ≥ 30bp.

[0065] The first gro...

Embodiment 2

[0091] The embodiment of the present invention is an example of using the identification method of mycobacterium tuberculosis and non-tuberculosis mycobacteria to identify target detection strains, please refer to Figure 1 to Figure 6 , Figure 16 to Figure 19 :

[0092] What needs to be explained first is that the target detection strains selected in this embodiment are as follows: Figure 5 and Image 6 As shown, the designed sequence information is as Figure 16 shown.

[0093] Homologous genes / sequences 16srRNA, 16s-23s rRNA interspace (ITS), mku, mce3B were selected.

[0094] Obtain the target detection strain, analyze the gene sequence information of the target detection strain, and determine the design coordinates.

[0095] For the detection of two different SNPs from A / G, design PCR primers and extension primers. Both ends of the upstream and downstream primers of the PCR primers need to add common sequences, namely ACGTTGGATG. The main purpose is to make the PCR...

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Abstract

The invention discloses a method for identifying mycobacterium piperculosis and nontuberculous mycobacterium. The method comprises the following steps: selecting homologous genes and sequences; acquiring a detected strain, and analyzing a gene sequence of the detected strain; designing a PCR primer and an extension primer; grouping the PCR primer and the extension primer to respectively obtain a PCR amplification primer mixed solution and a single-base extension primer mixed solution; carrying out PCR (Polymerase Chain Reaction) amplification on the detected strain to obtain a PCR product of a target site; carrying out dNTP elimination on the reaction system after PCR amplification to obtain a digested product; carrying out single-base extension reaction on the digested product to obtain an extended product; purifying the extension product to obtain a purified extension product; carrying out chip sample application and scanning through a nucleic acid detection mass spectrometer to obtain a detection result; subjecting the detection result to typing, outputting the result, obtaining a typing result, and identifying the strain through a mass spectrum peak; by adopting the method, the mycobacterium piperculosis complex group and the nontuberculous mycobacterium can be identified more accurately.

Description

technical field [0001] The invention relates to the technical field of gene detection, in particular to a method for identifying tuberculosis mycobacteria and non-tuberculosis mycobacteria. Background technique [0002] Tuberculosis is still a serious international public health problem, especially in developing countries. According to the statistics of the World Health Organization, 1 / 3 of the world's population is infected with Mycobacterium tuberculosis (MTB), and the infection of Non-tuberculous mycobacteria (NTM) is also on the rise. [0003] Both NTM and Mycobacterium tuberculosis complex (MTBC) can cause lung and other lesions, and the clinical symptoms are similar, making differential diagnosis difficult. [0004] At present, the methods commonly used in clinical identification of NTM are mainly divided into two categories. One is the traditional biochemical identification method, which is mainly based on bacterial growth rate, pigment production, drug tolerance, bi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/686C12Q1/04
CPCC12Q1/689C12Q1/686C12Q2533/101C12Q2565/627
Inventor 叶绍云蒋峻峰杨小娟刘雨
Owner 新起点生物科技苏州有限公司