A color-developing particle indicating microbial growth and its preparation method

A technology for indicating microorganisms and microorganisms, which is applied in the field of chromogenic particles and preparations indicating the growth of microorganisms, and can solve the problems of being relatively sensitive to temperature, humidity and light, poor indicating effect of positive bacteria, and slow growth of Mycobacterium tuberculosis.

Inactive Publication Date: 2011-11-30
丁元生
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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 ...

Method used

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  • A color-developing particle indicating microbial growth and its preparation method
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  • A color-developing particle indicating microbial growth and its preparation method

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preparation example Construction

[0059] The present invention also provides a method for preparing particles indicating the growth of microorganisms, the method comprising: mixing a redox indicator (optionally, a substance capable of proton transfer and / or a stabilizer) with the granulate to obtain an adherent Particles with (optionally also attached with a protonically capable substance and / or stabilizer) a redox indicator. The adding method and mixing sequence of the redox indicator, proton-transmitting substance and / or stabilizer are arbitrary, and the present invention has no special limitation.

[0060] The medium in which the oxidation-reduction indicator (optionally also includes proton-transferable substances and / or stabilizers) mixed with the granules can be water or an organic solvent. Described organic solvent is such as methanol, ethanol etc. The amount of water or organic solvent used is the minimum amount to ensure uniform mixing. Increasing the amount of solvent will not affect the effect, but...

specific Embodiment approach

[0063] 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.

[0064]As a preferred embodiment of the present invention, there is provided a specific method for preparing indicator particles indicating microbial growth, including: (1) adding a certain amount of tetrazolium salt (TTC, MTT, INT, TNBT, NBT, One of OTC, 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 gel particles are mixed evenly and dried in an oven to make color indicator particles, so that tetrazolium salts and the like are adsorbed on the surface of the silica gel particles.

[0065] The color indicator particle can be used as a general reagent. Before inoculation or...

Embodiment 1

[0071] Example 1, preparation of color indicating particles

[0072] 1. Reagent preparation

[0073] (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.

[0074] (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.

[0075] (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.

[0076] (4) Prepare 0.1M citric acid solution, aliquot after sterile filtration, and store at 4°C.

[0077] 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.

[0078] 3. Dry the mixture in an oven at 80°C, s...

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Abstract

The invention relates to a color developing particle indicating the growth of microorganisms and a preparation method. The indicator particles of the invention have good stability, high sensitivity and low toxicity to microorganisms.

Description

technical field [0001] The invention belongs to the field of microorganism detection; more specifically, the invention relates to a color-developing particle indicating the growth of microorganisms and a preparation method 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 mol...

Claims

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Application Information

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IPC IPC(8): G01N27/26C12Q1/70C12Q1/06
Inventor 孙永军
Owner 丁元生
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