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Method for screening mycobacterium tuberculosis multidrug-resistance protein

A technology for Mycobacterium tuberculosis and multi-drug resistance, which is applied in biochemical equipment and methods, microbial measurement/inspection, material separation, etc., and can solve problems such as comparative analysis of protein composition without seeing whole cells

Inactive Publication Date: 2007-04-25
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no comparative analysis of whole-cell constituent proteins of M.tb clinical multidrug-resistant strains

Method used

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  • Method for screening mycobacterium tuberculosis multidrug-resistance protein
  • Method for screening mycobacterium tuberculosis multidrug-resistance protein
  • Method for screening mycobacterium tuberculosis multidrug-resistance protein

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0038] The cultivation of embodiment 1 Mycobacterium tuberculosis and drug susceptibility test

[0039] Mycobacterium tuberculosis standard strain Mycobacterium tuberculosis H37Rv (ATCC93009), clinical multidrug-resistant strains (8 strains), and clinical all-sensitive strains (9 strains) were provided by the Tuberculosis Laboratory of Shanghai Pulmonary Hospital. The strains were inoculated into Middlebrook 7H9Broth (DIFCO, Bection-Dickinson) added with 10% ADC (DIFCO, 0.5% bovine serum albumin, 0.2% glucose, 140mmol / LNaCl) and 0.5% glycerol, and placed in a static incubator at 37°C. Place and cultivate for 15-20 days, when the bacterial concentration reaches to 1×10 8 ~2×10 8 or optical density (OD 600 ) was collected between 0.8 and 1.

[0040] Heat in a water bath at 80°C for 2 hours to kill bacteria. After centrifugation at 5000rpm at 4°C, the supernatant was discarded (the supernatant was first autoclaved at 121°C for 30 minutes and then discarded), and the bacteria ...

Embodiment 2-10

[0042] For specific steps of Examples 2-10, see the patent application "A Method for Screening Mycobacterium Tuberculosis Drug-Resistant Proteins" (Application No. 200610117294.1)

Embodiment 2

[0043] The extraction of embodiment 2 thalline protein

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Abstract

The invention belongs to the field of molecular biology of screening novel antituberculosis drugs target, vaccinum antigen and testing novel markers. The invention provides a method for screening the protein of Bacillus tuberculosis rifampicine-resistant: First, cultivating multi-drug-resistant strains; Secondly, separating the were isolated strains of multi-drug-resistant and sensitive strains of multi drug-resistant strains and protein from sensitive strains separately; Thirdly, determining the multi-drug-resistant protein by comparing the proteins of multi-drug-resistant strains and sensitive strains; Finally, separating and identifying the multi-drug-resistant protein. This invention provides a method which can accurately separate and identify multi-drug-resistant protein of Bacillus tuberculosis. The invention provides novel way for better understand of the multi-drug-resistant mechanism of Bacillus tuberculosis, rapid detecting of clinical multi-drug-resistant strains and the development of vaccine and drug target.

Description

technical field [0001] The invention belongs to the field of molecular biology screening of novel anti-tuberculosis drug targets, vaccine antigens and detection targets. Specifically, the invention provides a method for screening multidrug-resistant proteins in mycobacterium tuberculosis through proteome technology. The invention also provides the application of the method in the screening of drug targets, the identification of detection markers of multidrug-resistant strains, the preparation of vaccines and the like. Background technique [0002] Tuberculosis still poses a huge threat to mankind. Nearly 32% of the world's population has been infected with Mycobacterium tuberculosis. About 2.7 million people die from tuberculosis every year, an average of more than 7,000 people die every day. The World Health Organization declared tuberculosis a global health emergency in 1993. [0003] my country is one of the 22 countries with a high burden of tuberculosis in the world,...

Claims

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Application Information

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IPC IPC(8): C12Q1/04C12Q1/18G01N27/26G01N30/72G01N33/68
Inventor 王洪海王庆忠张鹭乐军张旻徐颖祝秉东王九龄郄亚卿
Owner FUDAN UNIV
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