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Large brill rhabdo virus toxic strain and its preparation method and application

A rhabdovirus and turbot technology, applied in the field of in vitro culture and identification, aquatic animal disease pathogens and isolation, can solve the problems of difficult control of fish infection experimental conditions, high cost, poor repeatability, etc.

Inactive Publication Date: 2004-06-02
INST OF AQUATIC LIFE ACAD SINICA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] Another object of the present invention also relates to a preparation method for preparing turbot rhabdovirus strain, which is simple and easy, and can amplify flounder rhabdovirus in a large amount in vitro, and solves the problem of turbot rhabdovirus in fish. The experimental conditions of body infection are difficult to control, the cost is high, and the repeatability is poor

Method used

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  • Large brill rhabdo virus toxic strain and its preparation method and application
  • Large brill rhabdo virus toxic strain and its preparation method and application
  • Large brill rhabdo virus toxic strain and its preparation method and application

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Embodiment Construction

[0020] All utensils, media, and reagents used for cell culture and virus amplification are required to be sterilized by autoclaving or filter sterilization in advance to keep them sterile. The preparation methods of some commonly used reagents are as follows:

[0021] Phosphate buffered saline (PBS, Ca-free ++ , Mg ++ ):

[0022] NaCl 8.00g

[0023] KCl0.20g

[0024] Na 2 HPO 4 2.31g

[0025] K H 2 PO 4 0.31g

[0026] Dissolve in 500ml of double distilled water, add 5ml of 0.4% phenol red solution, stir evenly, add double distilled water to 1000ml. 10 lbs sterilized for 15 minutes for use.

[0027] Double Antibody (Penicillin, Streptomycin) Solution

[0028] will be 10 6 Unit Penicillin G and 10 6 μg streptomycin was dissolved in 100ml sterilized three-distilled water to make 10 per ml penicillin 4 unit, streptomycin 10 4 μg.

[0029] TC119 Medium

[0030] Add the purchased TC199 medium powder according to the amount required in the...

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Abstract

The Scophthalmus maximus rhabdovirus, SMV0207, with preservation number of CCTCC No. V202006, is prepared through taking liver, kidney, heart and spleen tissues of suffering Scophthalmus maximus, crushing, adding PBS for homogenation, adding amphitolerant, freezing, thawing, centrifugating to obtain coarse virus extracting liquid, filtering, inoculation to CLC0207 cell for further culture at constant temperature. The said process is simple and practical, and the virus strain may be amplified in CLC0207 cell fast in the amplification potency up to 108 TCID50 / ml. The virus strain may be used in preparing sea water fish rhabdovirus cell engineering vaccine.

Description

technical field [0001] The invention relates to pathogens of aquatic animal diseases and their separation, in vitro cultivation and identification technology, in particular to the in vitro propagation, detection and diagnosis technology of marine cultured fish viruses. More specifically, it relates to a turbot rhabdovirus strain, a preparation method of a turbot rhabdovirus strain, and also relates to a biological method in identifying seawater fish rhabdoviruses and seawater fish bombs. The use of virus in the preparation of cell engineering vaccine. Background technique [0002] Turbot (Scophthalmus maximus, turbo) is a cold-water bottom-dwelling flounder marine fish native to Europe, and it is also a well-known marine cultured species in the local area. Turbot has a smooth, tender and delicious taste, less bone spurs, no fishy smell, and rich gelatin on the edge of the fins and under the skin, which is comparable to soft-shelled turtles and se...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/12C12N7/01C12Q1/70
Inventor 张奇亚李正秋桂建芳
Owner INST OF AQUATIC LIFE ACAD SINICA
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