DNA extraction method in pulse field gel electrophoresis (PFGE) method parting of mycobacterium tuberculosis (M. tb)

A technology of Mycobacterium tuberculosis and extraction method, which is applied in the field of DNA extraction based on Mycobacterium tuberculosis PFGE method typing, can solve the problems such as the difficulty of removing the cell wall, and achieve the effect of reducing experimental costs, reducing dosage, and clear gene fragments

Pending Publication Date: 2020-09-29
DALI UNIV
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the process of PFGE typing of Mycobacterium tuberculosis, it is difficult to remove the cell wall due to the particularity of M.tb's own cell structure, so the application of PFGE in M.tb genotyping is controversial among researchers. The main reason why there are few related reports on M.tb genotyping in recent years.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • DNA extraction method in pulse field gel electrophoresis (PFGE) method parting of mycobacterium tuberculosis (M. tb)
  • DNA extraction method in pulse field gel electrophoresis (PFGE) method parting of mycobacterium tuberculosis (M. tb)
  • DNA extraction method in pulse field gel electrophoresis (PFGE) method parting of mycobacterium tuberculosis (M. tb)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] 1. Subjects

[0048] Take sputum samples from 27 clinical pulmonary tuberculosis patients;

[0049] 2. Grouping

[0050] Divide 27 oral sputum samples into experimental group 1 and experimental group 2;

[0051] 3. Specimen processing

[0052] Take the mouth sputum specimens of pulmonary tuberculosis patients for acid treatment for 20 minutes, and neutralize with alkali;

[0053] 4. Cultivate

[0054] Inoculate in L-J (Roche) medium by partition marking method, and culture at 37°C;

[0055] 5. Purification

[0056] Observe every day, if there is colony growth, carry out: A. Observe the size, shape, edge, surface, color, etc. of the colony; B. Identification of acid-fast staining and identification medium; For biochemical reactions, if necessary, use the BacT / ALERT 3D 360 automatic culture instrument of the French Mérieux company and the matching BacT / ALERT MP Mycobacterium tuberculosis culture bottle for culture and identification; D. After the identification is c...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses a DNA extraction method in pulse field gel electrophoresis (PFGE) method parting of mycobacterium tuberculosis (M. tb). The DNA extraction method comprises the following steps:preparing a bacterial suspension from cultured M. tb in a bacterium grinding tube, performing inactivation, centrifugation and boiling water heating treatment on the bacterial suspension, then, adding a lysozyme, and placing the bacterial suspension in a plug mold to be solidified into a gel plug for later use; and carrying out cytolysis and water-bath oscillation on the gel plug repeatedly to obtain DNA of the M. tb. The method can conveniently remove cell walls of the M. tb and obtain the DNA of the M. tb; and besides, the method can reduce the cost and has higher promotion performance andpracticability.

Description

technical field [0001] The invention relates to the technical field of biological extraction, in particular to a DNA extraction method based on the PFGE typing of Mycobacterium tuberculosis. Background technique [0002] Tuberculosis is a major infectious disease causing adult mortality worldwide. With the development of molecular microbiology, many new methods for identifying microorganisms have been established and applied in recent years, but pulse field gel electrophoresis (PFGE) has always been a method of molecular epidemiology research on infectious diseases. important method of identification. The application of PFGE in the genotyping of Mycobacterium tuberculosis (M.tb) is helpful for the investigation of outbreaks of tuberculosis and the distinction between new infections and priming infections of old lesions. In the previous research, the researchers found that the resolution of PFGE in the genotyping of M.tb clinical isolates from pulmonary tuberculosis patient...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12N15/10
CPCC12N15/1003
Inventor 吴利先王国富聂恒王旭东郝贤斌李萍李丛哲
Owner DALI UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap