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Polyase chain reaction detecting method of Campylobacter jejuni

A technology of Campylobacter jejuni and a detection method, applied in the field of medical biology, can solve the problem of low specificity of Campylobacter jejuni and Campylobacter colon, and achieve the effects of increasing sensitivity and specificity and taking a long time to resolve.

Inactive Publication Date: 2009-01-28
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the PCR method reported at home and abroad has the defect of low specificity for the identification of Campylobacter jejuni and Campylobacter coli

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] 1. Isolation and cultivation of suspicious colonies: Take the chicken liver under sterile conditions, put it into a penicillin bottle with an appropriate amount of sterilized saline, filter it with a 0.65um microporous membrane, and inoculate it directly in the modified Camp-BAP selection culture cultured at 42°C in a microaerobic culture device.

[0018] 2. Pick suspicious colonies: Pick suspicious colonies from the colonies obtained from sample culture, among which: Type I suspicious colonies are non-hemolytic, off-white, brownish-yellow, flat, moist, shiny, translucent drop-shaped, raised and smooth , with irregular edges and a tendency to spread outward from the inoculation line; type II suspicious colonies are also hemolytic, round, shiny, translucent, and often scattered and raised single colonies (1-2 mm in diameter), with intact edges and hair Bright. The above-mentioned colonies can be further identified as suspicious colonies of Campylobacter jejuni.

[0019...

Embodiment 2

[0028] 1. Isolation and cultivation of suspicious colonies: under sterile conditions, take pig cecum and put it into a penicillin bottle with an appropriate amount of sterilized saline, filter it with a 0.65um microporous membrane, and inoculate it directly on the improved Camp-BAP selection medium Plates were cultured at 42°C in a microaerobic culture device.

[0029] 2. Pick suspicious colonies: Pick suspicious colonies from the colonies obtained from sample culture, among them: Type I suspicious colonies are not hemolytic, off-white, brownish yellow (may appear light red), flat, moist, shiny, and translucent in appearance Like water droplets, raised, smooth, with irregular edges and a tendency to spread outward from the inoculation line; type II suspicious colonies are also hemolytic, round, shiny, translucent, and often appear as scattered raised single colonies (with a diameter of 1 to 2mm), the edges are complete and shiny.

[0030] 3. Preparation of PCR template Pick a...

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Abstract

The invention relates to a detection method of the polymerase chain reaction of jejunum campylobacter, comprising the steps that micro-oxygen-needed separated cultivation is carried out to the jejunum campylobacter, suspicious colony is selected to be dissolved in a buffer solution specially used for dissolving the jejunum campylobacter, supernatant is taken out to be used as the template of PCR after boiling decentralization; then, a pair of specific primers are designed according to a conservative segment in a VS1 gene sequence of the jejunum campylobacter to carry out PCR reaction, the reaction product is observed by using agarose gel electrophoresis, and positive result is obtained if a 516bp strip appears. The method of the invention detects the jejunum campylobacter directly from the suspicious colony, the process of bacteria-increasing cultivation is eliminated, and the designed specific primers improve the sensitivity and the specificity of the PCR detection method, thus solving the difficulty of long detection time of the jejunum campylobacter by the conventional method, and providing a reliable and fast detection method for the detection of the jejunum campylobacter in livestock, fowl and the products thereof.

Description

technical field [0001] The invention relates to a detection method of campylobacter jejuni, a zoonotic pathogen, in particular to a polymerase chain reaction (PCR) detection method of campylobacter jejuni, which belongs to the field of medical biotechnology. Background technique [0002] Campylobacter jejuni (Campylobacter jejuni) is a kind of Campylobacter genus. It is a zoonotic pathogen that has received extensive attention at home and abroad in the past ten years. It can mainly cause acute enteritis and food poisoning in humans, and it is accompanied by reactive arthritis and hepatitis. , Reiter's disease and Guillain-Barré syndrome and other immune-impairing diseases. In veterinary medicine, it can cause abortion and infertility in livestock, mastitis, diarrhea in young livestock and poultry, and poultry hepatitis. The bacterium enters the environment through the feces of animal carriers or sick people, and can enter a dormant period called "viable but nonculturer-able...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04
CPCY02A50/30
Inventor 朱建国侯建军姜毅吴忠亮华修国
Owner SHANGHAI JIAO TONG UNIV
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