2-amino-2-deoxy-D-glucose and new application of hydrochloride, sulfate and myo-inositol thereof

A technology of glucose and hydrochloride, applied in the field of chemical compounds, can solve the problems of slow growth of Mycobacterium tuberculosis and achieve good application prospects

Inactive Publication Date: 2013-09-18
GANSU AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because the growth rate of Mycobacterium tuberculosis is very slow, it generally takes 28 days to form colonies visible to the naked eye in the commonly used Roche (Lowenstein-Jensen) solid medium. brings great challenges

Method used

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  • 2-amino-2-deoxy-D-glucose and new application of hydrochloride, sulfate and myo-inositol thereof
  • 2-amino-2-deoxy-D-glucose and new application of hydrochloride, sulfate and myo-inositol thereof
  • 2-amino-2-deoxy-D-glucose and new application of hydrochloride, sulfate and myo-inositol thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Preparation of control medium 7H9-S liquid medium:

[0023] Accurately weigh 0.47 g of 7H9, 0.2 ml of glycerin, and 0.05 ml of Tween-80 on an electronic balance, add 90 ml of ultrapure water, autoclave at 121°C for 10 minutes, cool to room temperature, and then add 10 ml of ADC nutrient solution to obtain Prepared 7H9-S liquid medium. The preparation of the ADC nutrient solution is as follows: Weigh 0.85 grams of sodium chloride, 5 grams of calf serum fraction V, 2 grams of glucose, and 0.003 grams of catalase, add 100ml of ultrapure water to fully dissolve, and then use 0.22um sterile The membrane was sterilized to obtain the prepared ADC nutrient solution.

Embodiment 2

[0025] Preparation of medium for active test group:

[0026]A certain amount of 2-amino-2-deoxy-D-glucose and its hydrochloride, sulfate and myo-inositol were accurately weighed with an electronic balance and put into 4 test tubes (the samples used in the test were purchased from Sangon Bioengineering Shanghai Co., Ltd., the purity of the four samples is 98%), respectively dissolved in sterile water, the concentration is 50 micromol / ml, respectively, sterilized with 0.22um sterile filter membrane and loaded into 4 Store in a sterile centrifuge tube. Then, the above four compound solutions prepared were diluted to 10 micromol / ml, 5 micromol / ml, 1 micromol / ml, and 0.1 micromol / ml with the prepared 7H9-S liquid medium respectively. Concentration gradient.

Embodiment 3

[0028] Growth-promoting activity assay:

[0029] Take out the attenuated Mycobacterium tuberculosis strain H37Ra preserved with glycerin from the -80°C low-temperature refrigerator, place it at 4°C for 30 minutes, and then dilute it with sterile water to a concentration of 1.0×10 8 CUF / ml, each 100ul was inoculated into 5ml of 7H9-S control medium and each test group medium test tube, cultured in a 37°C incubator for 5 days, read the absorbance value at OD600nm with a microplate reader, In order to detect the growth density of the H37Ra strain in each test group, and count the colonies of each test group on the 7H10 plate (purchased from BD Company of the United States, product number 262710) (wherein the attenuated strain of tuberculosis H37Ra was provided by Johns Hopkins, USA). Professor Ying Zhang from the Institute of Molecular Microbiology and Immunology of the University provided to this laboratory). The results show that the above four compounds have very significant ...

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Abstract

The invention discloses 2-amino-2-deoxy-D-glucose and an application of hydrochloride, sulfate and myo-inositol thereof in preparing a mycobacterium tuberculosis culture medium or a bacillus calmette guerin vaccine mycobacterium culture medium. The 2-amino-2-deoxy-D-glucose and the hydrochloride, sulfate and myo-inositol thereof serving as a growth factor of mycobacterium tuberculosis have good application prospects in developing a novel mycobacterium tuberculosis rapid culture medium, and can be applied to the rapid diagnosis of tuberculosis, type identification and drug sensitivity detection; and by adding one of the four compounds or the mixture of several ones of the four compounds into the culture medium, the flora density in the liquid culture medium is far higher than that of a blank control group, the macroscopic bacterial colony appears one week earlier in the solid culture medium than in the blank control group, and the bacterial plaque is obviously larger than that of the blank control group.

Description

[0001] technical field [0002] The present invention relates to the new application of compounds, in particular to the new application of 2-amino-2-deoxy-D-glucose and its hydrochloride, sulfate and myo-inositol. Background technique [0003] Tuberculosis is a highly contagious disease worldwide. One-third of the world's population is infected with Mycobacterium tuberculosis, and the number of deaths caused by tuberculosis is more than one million each year (Andreu N, Fletcher T, Krishnan N, Wiles S, Robertson BD.. J Antimicrob Chemother. 2012, 67: 404–414). Because the growth rate of Mycobacterium tuberculosis is very slow, it generally takes 28 days to form colonies visible to the naked eye in the commonly used Roche (Lowenstein-Jensen) solid medium. bring great challenges. In recent years, with the emergence of multidrug-resistant Mycobacterium tuberculosis (MDR-TB) and extensively drug-resistant Mycobacterium tuberculosis (XDR-TB) (Espinal M A, Laszlo A, Simonsen L, e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/04C12N1/20
Inventor 汪月张颖祝秉东李晶晶亢鸿飞
Owner GANSU AGRI UNIV
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