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Canine mycoplasma culture medium and preparation method thereof

A technology of Mycoplasma canis and liquid culture medium, applied in the field of microorganisms, can solve the problems of slow growth, high nutrient requirements of culture medium, slow progress of basic research, etc., and achieve the effect of high viable bacteria titer, rapid separation and analysis

Inactive Publication Date: 2019-12-31
陈永鑫
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Problems solved by technology

However, because Mycoplasma canis is very sensitive to the external environment, it has very high nutritional requirements for the medium, grows abnormally slowly, and is very difficult to separate. The conventional Mycoplasma medium is very poor for the isolation and cultivation of Mycoplasma canis, which leads to its basic research and clinical vaccines. and drug research and development are slow, so there is an urgent need to develop a medium suitable for the rapid and effective cultivation of Mycoplasma canis

Method used

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  • Canine mycoplasma culture medium and preparation method thereof
  • Canine mycoplasma culture medium and preparation method thereof

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Embodiment 1

[0016] Example 1 Mycoplasma canis high-efficiency medium and its preparation

[0017] The invention provides two mediums suitable for rapid and effective cultivation of Mycoplasma canis, which are respectively QG2 and QG5 mediums, each of which includes a liquid medium and a solid medium.

[0018] The formula of QG2 liquid medium is: 60%-64% of FM-4 basic culture medium, 10%-12% of yeast extract (30%), 5%-8% of calf serum, 1.0%-2.0% of dog serum, polypeptide T2 (4ng / ml) 1.0%-1.5%, L-asparagine (1g / ml) 0.02%-0.05%, NaCl (5g / ml) 0.12%-0.16%, penicillin (1×10 5 IU / ml) 1.2%, phenol red (1%) 0.003%, pH adjusted to 7.6-7.8. Preferably, the optimal QG2 liquid medium formula is: FM-4 basal culture medium 62%, yeast extract (30%) 10%, calf serum 6%, canine serum 1.5%, polypeptide T2 (4ng / ml) 1.2 %, L-asparagine (1g / ml) 0.04%, NaCl (5g / ml) 0.13%, penicillin (1×10 5 IU / ml) 1.2%, phenol red (1%) 0.003%, pH adjusted to 7.6-7.8.

[0019] The formula of QG5 liquid medium is: FM-4 basic c...

Embodiment 2

[0024] Example 2 Effectiveness of Mycoplasma canis high-efficiency medium

[0025] Then test the effect of QG medium on the cultivation of Mycoplasma canis. In a pet breeding base in Fuzhou City, Fujian Province, anticoagulant venous blood was collected from dogs with suspected mycoplasma disease (polymorphic spheroid microorganisms can be seen on blood smear Giemsa staining), and 0.1ml of each was added to 100ml of QG liquid culture medium (including QG2 and QG5) and three conventional mycoplasma liquid media (including FM-4, PPLO and SP4 medium, as a control) (the first inoculation), placed at 37°C and 6% CO 2 Cultivate under the same conditions until the color of the medium changes from red to yellow, and then take 10ml of the culture solution and add it to a new 90ml of the respective liquid medium. Cultivate under the same conditions until the medium changes color (the second inoculation), and repeat the transfer once (the second inoculation). Three inoculations), and fi...

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Abstract

The present invention provides a new and high-efficient canine mycoplasma culture medium QG5. The culture medium contains canine serum, polypeptides T5 as shown in SEQ ID:2, L-asparaginate and NaCl based on mycoplasma conventional culture medium FM-4. The canine mycoplasma can achieve rapid growth and proliferation in the QG5 culture medium and also reaches a very high viable micro-organism titer,which helps rapid detection and diagnosis of the canine mycoplasma, helps developments of canine mycoplasma vaccines and therapeutic drugs and helps to study mechanisms of the canine mycoplasma disease.

Description

technical field [0001] The invention relates to the field of microorganisms, in particular to culture medium for mycoplasma canis and application thereof. Background technique [0002] Mycoplasma is currently known as the prokaryotic microorganism with the smallest size and the simplest structure that can live independently. It has no cell wall, no flagella, no movement, Gram staining is negative, but it is not easy to color, and it shows polymorphic spheroids in Giemsa stained slides . A variety of mycoplasma can infect dogs. Mycoplasma canis mainly causes canine respiratory diseases, genitourinary tract diseases, anemia, arthritis and colitis, etc. For example, dogs are susceptible to mycoplasma haemophilus and cause persistent fever, runny nose, and loss of appetite. In severe cases, acute clinical symptoms such as vomiting and diarrhea will occur, and severe anemia or hemolytic anemia will occur, and eventually the patient will die of blood in the stool. Therefore, myc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12R1/35
CPCC12N1/20
Inventor 陈永鑫黄明水
Owner 陈永鑫
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