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Microaerobic isolated culture method of campylobacter jejuni

A technology for isolation and culture of Campylobacter jejuni, which is applied in the field of medical microorganisms, can solve the problems of low detection rate, difficult to obtain materials, and high requirements for experimental conditions, and achieves the effects of high detection rate, large number of colonies, and good culture effect.

Inactive Publication Date: 2009-03-25
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above methods have problems such as high requirements on experimental conditions, or difficult to obtain materials, or low detection rate.

Method used

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  • Microaerobic isolated culture method of campylobacter jejuni

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] 1. Material collection: Take chicken liver and put it into a penicillin bottle with an appropriate amount of sterilized saline, and cut the liver aseptically to about 0.8~1.0cm 3 , 4C save for future use.

[0018] 2. Isolation and culture of Campylobacter jejuni: using direct streaking plate culture method, grind the collected chicken liver, first filter it with a 0.65um microporous membrane, and then take the filtrate and inoculate it on the modified Camp-BAP medium. ; Place the plate that has been inoculated with bacteria in a 3000mL grinder dryer. Coat the cylinder head and the cylinder mouth with petroleum jelly, add coke gallic acid and anhydrous sodium carbonate at the amount of 2.6g per 1L volume (1:1 ratio), mix well, put a small piece of candle in the center of the cylinder, cover the cylinder cover. After burning the candle for about 0.5 to 1 min, it will extinguish by itself, and place it in the incubator together with the cylinder for 24 hours at 42°C.

[0019] ...

Embodiment 2

[0030] 1. Material collection: Take pig liver and put it into a penicillin bottle with an appropriate amount of sterilized physiological saline, and cut the liver about 0.8~1.0cm aseptically 3 , 4C save for future use.

[0031] 2. Separation and culture of Campylobacter jejuni: using direct streaking plate culture method, grind the collected pig liver, first filter it with a 0.65um microporous membrane, and then take the filtrate and inoculate it on the modified Camp-BAP medium ; Place the plate that has been inoculated with bacteria in a 3000mL grinder dryer. Coat the cylinder head and the cylinder mouth with petroleum jelly, add coke gallic acid and anhydrous sodium carbonate according to the amount of 2.4g per 1L volume (1:1 ratio), mix evenly, put a small piece of lighted candle in the center of the cylinder, cover the cylinder cover. After burning the candle for about 0.5 to 1 min, it will extinguish by itself, and place it together with the container in an incubator at 42°C ...

Embodiment 3

[0037] 1. Material collection: Take chicken liver and put it into a penicillin bottle with an appropriate amount of sterilized saline, and cut the liver aseptically to about 0.8~1.0cm 3 , 4C save for future use.

[0038] 2. Isolation and culture of Campylobacter jejuni: using direct streaking plate culture method, grind the collected chicken liver, first filter it with a 0.65um microporous membrane, and then take the filtrate and inoculate it on the modified Camp-BAP medium. ; Place the plate that has been inoculated with bacteria in a 3000mL grinder dryer. Coat the cylinder head and the cylinder mouth with petroleum jelly, add the coke gallic acid and anhydrous sodium carbonate at the amount of 3.0g per 1L volume (1:1 ratio), mix well, put a small piece of candle in the center of the cylinder, cover the cylinder cover. After burning the candle for about 0.5 to 1 min, it will extinguish by itself, and place it in the incubator together with the cylinder for 36 hours at 42°C.

[00...

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Abstract

The invention relates to a microaerophilic separation culture method of Campylobacter jejuni, which adopts the combination of a general candle-cylinder method and a pyrogallic acid method to create a microaerophilic environment. A directive streak cultivation method is adopted against the separation culture of the Campylobacter jejuni, a small amount of animal tissue detection samples are picked and directly inoculated on the selected culture medium by using connection annulus, the concentration of oxygen in the air is reduced by the candle-cylinder method with the combination of pyrogallic acid and anhydrous sodium carbonate mixture, an alkaline environment is formed by using anhydrous sodium carbonate so as to lead the pyrogallic acid to react with oxygen for absorbing the oxygen, therefore the culture environment meets the microaerophilic conditions required by the Campylobacter jejuni. Typical colonies obtained from separation culture are picked for PCR identification so as to further determine the Campylobacter jejuni. The microaerophilic separation culture method of Campylobacter jejuni has the advantages of low cost, simple operation, high detectable rate, good culture efficiency and the like, thus improving the separation culture efficiency of the Campylobacter jejuni and resolving the difficulty for cultivating the Campylobacter jejuni under microaerophilic condition.

Description

Technical field [0001] The invention relates to a method for separating and cultivating Campylobacter jejuni, in particular to a method for microaerobic separation and culturing of Campylobacter jejuni, a pathogen of zoonotic diseases. It belongs to the field of medical microbiology technology. Background technique [0002] Campylobader jejuni (Campylobader jejuni) is a species of Campylobacter. Campylobacter jejuni is widespread in nature and seriously threatens the health of humans and animals. It is a zoonotic pathogen that has been widely valued by scholars at home and abroad in the past ten years. Campylobacter jejuni infection has now become one of the most common causes of acute gastroenteritis worldwide. In developing countries, Campylobacter jejuni is an important cause of death from diarrhea in children, and it is also the most common cause of travellers’ diarrhea in developing countries. A serious complication is the acute demyelinating disease of the peripheral nervou...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12Q1/68C12Q1/04C12R1/01
Inventor 朱建国侯建军姜毅吴忠亮华修国
Owner SHANGHAI JIAO TONG UNIV
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