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Indicator tube for indicating microbe growth and application thereof

A technology for indicating microorganisms and indicating tubes, which is applied to an indicating tube indicating the growth of microorganisms and its application field, which can solve the problems of inability to reduce the color of the indicator, easy degradation, and reducing the indicating ability of tetrazolium salts.

Inactive Publication Date: 2012-02-08
新疆维吾尔自治区胸科医院 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] 1. Tetrazolium salt is unstable, sensitive to temperature, humidity and light, and easy to degrade, especially in aqueous solution, it is easy to generate colored products, and its storage solution should not exceed one month even if it is stored in a low temperature (-20°C) environment
The degradation or discoloration of tetrazolium salt greatly reduces the sensitivity of detection. Therefore, it is generally recommended that the tetrazolium salt solution be freshly prepared and used up as soon as possible, which severely limits the use of tetrazolium salt, especially when a certain shelf life is required. Applications in culture, identification, drug sensitivity reagents or products
[0012] 2. In fact, tetrazolium substances are toxic to the growth of bacteria, large or small. Studies have shown that when the TTC content of tetrazolium salt is 0.005%, the red indicator effect of Gram-negative bacteria is the best, and it has the best effect on bacteria. It is also small, but it has a strong inhibitory effect on most Gram-positive bacteria (such as mycobacteria, etc.). If the concentration is lowered, it will not be effective in indicating positive bacteria.
For example, if TTC is used as an indicator for food microbial colony counting determination, unqualified food with excessive microorganisms may be misjudged as qualified food
[0013] 3. It cannot be added to the medium of some slow-growing bacteria (such as mycobacteria, fungi, etc.) in advance to indicate their growth, especially Mycobacterium tuberculosis
[0014] It should be said that tetrazoles have been widely used in cell culture and the cultivation, identification and drug sensitivity of fast-growing bacteria, but they are limited in the use of mycobacteria, especially Mycobacterium tuberculosis. The reasons are: (1) tuberculosis classification The nutritional conditions required for the growth of mycobacteria are very harsh, and they are more sensitive to tetrazole substances. If the added amount is large, it will inhibit its growth, and if it is small, it will not see the change of color; (2) Mycobacterium tuberculosis grows extremely slowly (breeding one generation The cycle takes 18-20 hours), on the one hand, the energy metabolism is very low, and when there is only a small amount of bacteria in the initial growth period, the indicator cannot be restored to color at all.
On the other hand, the tetrazolium salt in the culture medium is easily degraded when cultured at 37°C for several days, more than ten days or even twenty days, which reduces the indicator ability of the tetrazolium salt.

Method used

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  • Indicator tube for indicating microbe growth and application thereof
  • Indicator tube for indicating microbe growth and application thereof
  • Indicator tube for indicating microbe growth and application thereof

Examples

Experimental program
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Effect test

specific Embodiment approach

[0080] As a specific embodiment of the present invention, a tetrazolium salt redox indicator is adsorbed on silica gel particles to prepare stable and low-toxic color indicator particles.

[0081] The amount of indicator particles in the growth indicator tube should be sufficient to develop color. Generally, it can be determined according to the container and the amount of sample (or sample solution, or culture solution) added, for example, 0.02-0.5g / tube (2ml-5ml).

[0082] As a preferred embodiment of the present invention, a method for preparing an indicator tube for specific microbial growth is provided, including: (1) adding a certain amount of tetrazolium salt (TTC, MTT, INT, TNBT, NBT, OTC, One of STC, NTC, PTC, MTC, etc.) and a certain amount of substances that stabilize tetrazolium salts (boric acid, citric acid or succinic acid) and a certain amount of proton-transferable substances PMS or PES and pre-heated at 130°C After activating for 2 hours, the cooled silica g...

Embodiment 1

[0093] Embodiment 1, the empty pipe that does not load into particle

[0094] Empty tubes not loaded with pellets such as figure 1 shown.

[0095] One kind of empty pipe is the case where the bottom of the pipe is a round bottom, such as figure 1 Middle left image. Among them, 1 is the tube body; 2 is the screw cap; 3 is the bottom of the bottle.

[0096] Another kind of empty pipe is the situation where the bottom of the pipe is narrowed, such as figure 1 Middle right image. Among them, 1 is the tube body; 2 is the screw cap; 4 is the bottom of the bottle.

Embodiment 2

[0097] Example 2, preparation of color indicator particles and growth indicator tube

[0098] 1. Reagent preparation

[0099] (1) Weigh 30 g of 200-mesh silica gel (purchased from Shanghai Shangbang Industrial Co., Ltd.), place it in an oven at 120°C for 2 hours, seal it and cool it down.

[0100] (2) Weigh 40 mg of TTC and dissolve it in 40 ml of distilled water (1 mg / ml), filter it aseptically, aliquot it, and store it in the dark at 4°C.

[0101] (3) Weigh 40 mg of PMS and dissolve it in 40 ml of distilled water (1 mg / ml), filter it aseptically, aliquot it, and store it in the dark at 4°C.

[0102] (4) Prepare 0.1M citric acid solution, aliquot after sterile filtration, and store at 4°C.

[0103] 2. Take 8ml of 0.1M citric acid solution in a sterile environment and add it to 40ml 1mg / ml tetrazolium salt (TTC) solution, then add 0.2ml 1mg / ml PMS, mix and pour into 30g activated silica gel particles, Stir with a sterilized glass rod until uniform.

[0104] 3. Dry the mixt...

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Abstract

The invention relates to an indicator tube for indicating microbe growth and application thereof. As a general container, the indicator tube of the invention employs a drying method to conduct coloration for indicating storage and transportation of particles; so that indicating particles are maintained at a drying state be for usage, and influence of humidity on a redox indicator (e.g. tetrazole salt) is avoided, and the redox indicator is not susceptible to degradation or color changing. The invention also provides a kit for detecting microbe growth. The indicator tube of the invention is easily used, has good versatility, high sensitivity and low toxicity.

Description

technical field [0001] The invention belongs to the field of microorganism detection; more specifically, the invention relates to an indicator tube for indicating the growth of microorganisms and an application thereof. Background technique [0002] Clinically, the substrate for pathogenic bacteria to obtain energy is mainly sugar, which releases energy through sugar oxidation or fermentation, and stores energy in the form of high-energy phosphate bonds (ADP, ATP). The types of bacterial biooxidation are divided into respiration and fermentation. During the biochemical process, the hydrogen removed from the nutrients of bacteria (such as sugar) by dehydrogenase needs to be transported by a series of intermediate hydrogen transfer bodies (such as coenzyme I, coenzyme II, flavoprotein, etc.). Finally, the hydrogen is given to the hydrogen acceptor. The biological oxidation process with inorganic substances as hydrogen acceptors is called respiration, in which molecular oxyge...

Claims

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Application Information

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IPC IPC(8): C12M1/34C12M1/24C12Q1/06C12Q1/04
CPCC12M23/08C12M41/36C12M25/16
Inventor 綦迎成李君莲张子羿丁元生
Owner 新疆维吾尔自治区胸科医院
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