Fusobacterium necrophorum selective medium

A technology for selecting culture medium and Fusobacterium necrosis, applied in microorganism-based methods, microbial determination/inspection, microorganisms, etc., can solve the problems of high cost, complicated operation, lack of anaerobic inspection equipment, etc., and achieve reliable and reliable detection. The effect of high separation rate

Inactive Publication Date: 2015-09-09
鲁莉
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the detection method of Fusobacterium necroptosis mainly adopts the molecular biology detection method based on polymerase chain reaction (PCR). Instruments and equipment are expensive, complicated to operate, and difficult to implement in grassroots laboratories
CDC anaerobic bacteria agar is commonly used in laboratories to cultivate Fusobacterium necrosis. Since most laboratory departments of primary hospitals lack anaerobic testing equipment, most of them use "anaerobic tank culture method". Put the flat plate or liquid medium test tube into the dry tank for cultivation, exhaust the oxygen in the tank by physical and chemical methods, and create an anaerobic environment
The oxygen depletion of the anaerobic cylinder method takes time, and Fusobacterium necroptosis is an obligate anaerobic bacillus, so this culture technique has the following defects: ① Fusobacterium necrosis grows slowly on the CDC anaerobic bacteria agar medium in the anaerobic cylinder , the positive detection rate is low, and a more accurate identification can be made in about a week; ②Because of the non-absolute anaerobic condition, aerobic bacteria and facultative anaerobic bacteria in throat secretion specimens have living conditions, and the separation ability of the medium is poor

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033]Example 1, the formula for preparing 1000ml medium contains: tryptone 10.0g; glucose 5.0g; yeast powder 5.0g; sodium chloride 5.0g; agar 15.0g; Potassium magnesium salt 3g; γ-aminobutyric acid 0.1g; sterile heat deformed calf serum 100ml; distilled water added to 1000ml. Preparation method: Weigh tryptone; glucose; yeast powder; sodium chloride; agar; ox bile salt; ethylenediaminetetraacetic acid dipotassium magnesium salt; Sterilize for 15 minutes, cool to 45°C, add sterile heat-deformed calf serum and mix evenly, dilute to 1000ml with sterilized distilled water and dispense.

[0034] The preparation method of the sterile heat-deformed calf serum is as follows: heat the sterile calf serum at 80°C for 10 minutes, add sterilized trypsin at 45°C for 90 minutes, and the mass ratio of enzyme to substrate is 1:150. The preparation method of the sterile heat-deformed calf serum in other examples is the same as that in Example 1.

Embodiment 2

[0035] Example 2, the formula for preparing 1000ml medium contains: tryptone 8.0g; glucose 10.0g; yeast powder 8.0g; sodium chloride 5.0g; agar 15.0g; Potassium magnesium salt 2.5g; γ-aminobutyric acid 0.3g; valine 0.1g; sterile heat deformed calf serum 50ml; distilled water added to 1000ml. Preparation method: Weigh tryptone; glucose; yeast powder; sodium chloride; agar; ox bile salt; dipotassium magnesium edetate; γ-aminobutyric acid; Homogenize, sterilize at 121°C for 15 minutes, cool to 45°C, add sterile heat-deformed calf serum, mix evenly, dilute to 1000ml with sterilized distilled water, and dispense.

Embodiment 3

[0036] Example 3, the formula for preparing 1000ml medium contains: tryptone 10.0g; glucose 8.0g; yeast powder 10.0g; sodium chloride 5.0g; agar 20.0g; bovine bile salt 0.75g; Potassium magnesium salt 3g; γ-aminobutyric acid 0.5g; valine 0.5g; L-homoarginine 0.05g; sterile heat deformed calf serum 80ml; distilled water added to 1000ml. Preparation method: weigh tryptone; glucose; yeast powder; sodium chloride; agar; ox bile salt; dipotassium magnesium EDTA; γ-aminobutyric acid; valine; , dissolved in distilled water, mixed evenly, sterilized at 121°C for 15 minutes, cooled to 45°C, added with sterile heat-deformed calf serum, mixed evenly, distilled to 1000ml in sterilized distilled water, and dispensed.

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Abstract

The invention discloses a fusobacterium necrophorum selective medium and a preparation method thereof and belongs to the field of microculture examination. The fusobacterium necrophorum selective medium is characterized in that 1000 ml of culture medium comprises casein tryptone, glucose, yeast powder, sodium chloride, agar, bile salt, ethylene diamine tetraacetic acid dipotassium magnesium salt, gamma-aminobutyric acid, sterile thermal-deformation calf serum and the balance of distilled water. Compared with the prior art, the fusobacterium necrophorum selective medium has the advantages of reliability in examination and high separation rate of fusobacterium necrophorum.

Description

technical field [0001] The invention relates to the field of culture of test microorganisms, in particular to a selective culture medium for fusobacterium necrosis. Background technique [0002] Microbial culture is a technology that uses artificial methods to grow and reproduce microorganisms, and selective medium is a medium designed to promote or inhibit certain types of organisms (such as cells or bacteria, etc.), using this medium can Separation of desired microorganisms from confounding microorganisms. Fusobacterium necrophorum (Fn) is a Gram-negative, strictly anaerobic bacterium of the genus Fusobacterium. Pathogenic Fn strains produce a variety of independent factors, such as leukotoxin (Lkt), hemolysin (hemolysin), hemagglutinin (hemagglutinin) and endotoxin (endotoxin), which can cause multiple mammals (including humans) and Necrobacillosis of poultry. Fusobacterium necroptosis is the main and secondary pathogen of various purulent necrotic infectious diseases,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/04C12R1/01
Inventor 鲁莉蒋欣雷志平董礼艳
Owner 鲁莉
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