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Preparation method of detecting system for minimal inhibitory concentration of mycobacteria

A minimum inhibitory concentration, mycobacteria technology, applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve problems such as complex operation, biological safety issues, increased contamination probability, etc., to achieve simple detection system and easy operation Simplified, low-pollution effect

Inactive Publication Date: 2012-07-11
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The currently used micro-color method is to first cultivate the bacterial solution for a period of time, then open the cover of the culture plate, add the display reagent, and then carry out a short-term color development, which is not only complicated to operate but also increases the probability of contamination biosafety issues

Method used

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  • Preparation method of detecting system for minimal inhibitory concentration of mycobacteria
  • Preparation method of detecting system for minimal inhibitory concentration of mycobacteria
  • Preparation method of detecting system for minimal inhibitory concentration of mycobacteria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Embodiment 1: 1. Select a sterile 96 microwell plate for use.

[0056] 2. Preparation of Middle brook 7H9 liquid medium: Take 4.9 grams of Middle brook 7H9 medium dry powder, add it to a reagent bottle containing 900ml of distilled water, fully dissolve, and sterilize at 121°C for 30 minutes. After the medium was cooled to room temperature, the nutritional supplement OADC (0.1% casein peptone, 0.5% glycerol, 10% oleic acid, bovine serum albumin, glucose and catalase) was added in a ratio of 10:1. Store at -4°C for later use.

[0057] 3, the preparation of bacterium liquid: after cultivating Mycobacterium tuberculosis clinical isolate A in Roche's culture medium (commercially available) for two weeks, get some bacterial colonies and resuspend with liquid culture medium, add 6-7 3-4mm glass beads in Grind the bacteria on a vortex shaker for 5-10 minutes, and after standing for 10 minutes, take the uniform suspension in the upper layer and use the liquid medium to compare...

Embodiment 2

[0065] 1. Select eight 1.5ml sterile centrifuge tubes for use.

[0066] 2. Preparation of Middle brook 7H9 liquid medium: Take 4.9 grams of Middle brook 7H9 medium dry powder, add it to a reagent bottle containing 900ml of distilled water, fully dissolve, and sterilize at 121°C for 30 minutes. After the medium was cooled to room temperature, the nutritional supplement OADC (0.1% casein peptone, 0.5% glycerol, 10% oleic acid, bovine serum albumin, glucose and catalase) was added in a ratio of 10:1. Store at -4°C for later use.

[0067] 3, the preparation of bacterium liquid: after cultivating Mycobacterium tuberculosis clinical isolate A in Roche's culture medium (commercially available) for two weeks, get some bacterial colonies and resuspend with liquid culture medium, add 6-7 3-4mm glass beads in Grind the bacteria on a vortex shaker for 5-10 minutes, and after standing for 10 minutes, take the uniform suspension in the upper layer and use the liquid medium to compare the t...

Embodiment 3

[0075] 1. Choose a sterile 96 microwell plate for use.

[0076] 2. Preparation of Middle brook 7H9 liquid medium: Take 4.9 grams of Middle brook 7H9 medium dry powder, add it to a reagent bottle containing 900ml of distilled water, fully dissolve, and sterilize at 121°C for 30 minutes. After the medium was cooled to room temperature, the nutritional supplement OADC (0.1% casein peptone, 0.5% glycerol, 10% oleic acid, bovine serum albumin, glucose and catalase) was added in a ratio of 10:1. Store at -4°C for later use.

[0077] 3, the preparation of bacterium liquid: after cultivating Mycobacterium tuberculosis clinical isolate A in Roche's culture medium (commercially available) for two weeks, get some bacterial colonies and resuspend with liquid culture medium, add 6-7 3-4mm glass beads in Grind the bacteria on a vortex shaker for 5-10 minutes, and after standing for 10 minutes, take the uniform suspension in the upper layer and use the liquid medium to compare the turbidity...

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Abstract

The invention provides a preparation method of a detecting system for minimal inhibitory concentration of mycobacteria, which includes the following steps: a prepared liquid nutrient medium containing medicine and a reacting system with certain volume of mycobacteria liquid and a color developing agent are co-cultured at certain temperature for a period of time, color-development detecting is conducted by adopting a certain detecting method, and the minimal inhibitory concentration of medicine is determined according to color development detecting results. Compared with the prior art, the preparation method co-cultures the liquid nutrient medium containing the medicine, the mycobacteria liquid, the color developing agent and intermediate electron acceptor, a step in the prior art that the color developing agent is added by opening a cover after a period of time of culture is omitted, operation is simplified, and operation pollution and biological safety risk are reduced. The detecting system is simple, safe, low in pollution and capable of being seen through eyes, and enables users to interpret the minimal inhibitory concentration of mycobacteria medicine fast.

Description

technical field [0001] The present invention relates to the detection of minimum inhibitory concentration (MIC) of mycobacteria or the field of antibacterial drug screening, in particular to a method for preparing a minimum inhibitory concentration (MIC) detection system of mycobacteria. The detection system is applicable to Drug susceptibility detection of mycobacteria, actinomycetes, fungi, cells, etc. Background technique [0002] So far, tuberculosis is still one of the most common infectious diseases in the world. Although the overall incidence of tuberculosis worldwide has begun to decline, drug resistance is becoming more and more serious. According to the World Health Organization (WHO), there were 8.8 million tuberculosis patients and 1.1 million deaths in the world in 2010. Incomplete treatment programs and management measures have led to a gradual increase in the rate and degree of drug resistance of tuberculosis. In 2010, at least 5% of new or relapsed cases bel...

Claims

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Application Information

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IPC IPC(8): C12Q1/18
Inventor 张舒林孙战强余晓丽张朝宝王洪海
Owner SHANGHAI JIAO TONG UNIV
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