Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Technical method for identifying human mycobacterium tuberculosis, mycobacterium bovis and bacillus calmette-guerin (BCG)

A technology for Mycobacterium tuberculosis and Mycobacterium bovis is applied in the field of biomedicine, and can solve the problems of deletion and inability to distinguish Mycobacterium tuberculosis complex Mycobacterium

Pending Publication Date: 2020-05-15
NANJING UNIV
View PDF2 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, 16S rRNA can identify multiple mycobacteria to species, but cannot distinguish the Mycobacterium tuberculosis complex from other mycobacteria
IS6110 is a multi-copy sequence with high sensitivity and can detect and identify the Mycobacterium tuberculosis complex, but it has been found to be missing in some MTB strains

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
  • Technical method for identifying human mycobacterium tuberculosis, mycobacterium bovis and bacillus calmette-guerin (BCG)
  • Technical method for identifying human mycobacterium tuberculosis, mycobacterium bovis and bacillus calmette-guerin (BCG)
  • Technical method for identifying human mycobacterium tuberculosis, mycobacterium bovis and bacillus calmette-guerin (BCG)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Multiple PCR detection of embodiment 1 human mycobacterium tuberculosis, mycobacterium bovis and BCG

[0034] (1) Preparation of samples

[0035] Using the bacteria shown below, DNA was extracted and purified by the following method to obtain a DNA sample.

[0036] a) Mycobacterium tuberculosis H37Ra strain

[0037] b) Mycobacterium tuberculosis BCG strain (BCG)

[0038] c) Mycobacterium bovis No. 552 strain

[0039] d) No. 739 strain of non-tuberculous mycobacteria;

[0040] e) Listeria strains

[0041] Specific steps are as follows:

[0042] Listeria was cultured in LB medium as a control. The extracted DNA was dissolved in sterile water, and the concentration of the obtained DNA was measured using NanoDrop 2000.

[0043] The modified CTAB method was used to extract the total DNA of Mycobacterium tuberculosis, and the specific steps were as follows:

[0044] 1. Use an inoculation loop to scrape a certain amount of bacteria into a centrifuge tube (400 μL TE buf...

Embodiment 2

[0068] Example 2 Preparation of strains to be tested

[0069] There are two methods for collecting the strains to be tested used in the detection method. The first way is to collect the sputum of clinically suspected tuberculosis patients, heat and boil the collected sputum at 100°C for 10 minutes, then centrifuge it at a high speed of 12000rpm / min for 10 minutes, and finally take the supernatant as a PCR template for use . In the second way, if the amount of bacteria collected in sputum is too small to meet the PCR detection standard, the modified Roche medium is used for amplification culture, and the cultured strains are subjected to the following operations.

[0070] The modified Roche medium is mostly used for the isolation and culture of mycobacteria, the preservation of strains and the identification of strains. Its culture of mycobacteria is accurate and reliable, and its specificity is high. Roche's medium contains basic substances, nutrients and bacteriostatic age...

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 provides a technical method for identifying human mycobacterium tuberculosis, mycobacterium bovis and bacillus calmette-guerin (BCG). Five target genes, i.e., 16S rRNA, IS6110, Rv2659c,ESAT6 and Rv1510 are selected for multiple PCR amplification, the mycobacterium tuberculosis can be rapidly and accurately identified, and interference caused by the mycobacterium bovis and BCG low-virulent strains (BCG) is effectively removed.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and specifically relates to a technical method for identifying human mycobacterium tuberculosis, mycobacterium bovis and BCG, which can quickly and effectively identify human mycobacterium tuberculosis. Background technique [0002] Tuberculosis is an infectious disease that seriously endangers the health of humans and animal husbandry. According to the estimates of the World Health Organization (WHO), there are currently 2 billion people infected with Mycobacterium tuberculosis (MTB) in the world, with an average of 1 new infection every 1 second, and 8 million to 10 million new tuberculosis patients every year , 3 million people died of tuberculosis, the infectious disease with the highest mortality rate. The pathogen that causes human and animal tuberculosis infection is Mycobacterium tuberculosis, which includes Mycobacterium tuberculosis complex (Mycobacterium tuberculosis complex, MTC)...

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
Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/686C12Q1/04C12R1/32
CPCC12Q1/689C12Q1/686C12Q2600/16C12Q2565/125C12Q2537/143
Inventor 王宏伟蒲文渊赵晨施旭东张侠苏忠兰
Owner NANJING UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com