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Lefteye flounder lymph tumour virus toxic strain, its preparation method and application

A technique for lymphocyst virus and flounder, which is applied to the flounder lymphocyst virus strain and the fields of preparation and application thereof, can solve the problems such as the flounder lymphocyst virus has not yet been established, the time required is long, the in vitro culture is difficult, and the like. The effect of manual adjustment and control, reduced detection cost, and easy experimental conditions

Inactive Publication Date: 2004-06-02
INST OF AQUATIC LIFE ACAD SINICA
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Problems solved by technology

[0004] Although the isolation and cultivation of fish lymphocyst virus began in the 1960s [Wolf et al, 1966, Science151: 1004-1005] and lasted about 40 years, the in vitro cultivation of this virus strain was very difficult, and only LCDV was observed Causes lesions in a small number of marine fish cells, and takes a long time, mostly over a week [Walker et al, 1980, J Gen. Virol.51: 385-395; Perez-Prieto et al, 1999, Dis Aquar Org 35 : 149-153; Chang et al, 2001, Aquaculture 192: 133-145]
So far, no economical and practical biological detection method for flounder lymphocyst virus has been established at home and abroad

Method used

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  • Lefteye flounder lymph tumour virus toxic strain, its preparation method and application
  • Lefteye flounder lymph tumour virus toxic strain, its preparation method and application
  • Lefteye flounder lymph tumour virus toxic strain, its preparation method and application

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Embodiment Construction

[0029] 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:

[0030] Phosphate buffered saline (PBS, Ca-free ++ , Mg ++ ):

[0031] NaCl 8.00g

[0032] KCl0.20g

[0033] Na 2 HPO 4 2.31g

[0034] K H 2 PO 4 0.31g

[0035] 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.

[0036] Double Antibody (Penicillin, Streptomycin) Solution

[0037] 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.

[0038] TC119 Medium

[0039] Add the purchased TC199 medium powder according to the amount required in the instr...

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Abstract

Paralichthys lymphocystis virus strain, LCDV011126, with the preservation number of CCTCC No. V202007, is prepared through taking Paralichthys tumor tissue of lymphocystis virus, crushing, adding phosphoric acid buffering liquid for homogenation, adding amphitolerant, freezing, thawing, centrifugating to obtain coarse virus extracting liquid, filtering, inoculation to fresh water fish cell for further culture at constant temperature, observing cell virus, collecting cell, fixing, superthin slicing, and dyeing for observation. The process is simple, easy to regulate and control and low in cost, and may be used in amplifying great amount of lymphocystis virus and providing required virus material.

Description

technical field [0001] The present invention relates to fresh and seawater fish disease pathogens and separation, amplification and identification technology, in particular to the in vitro propagation, detection and diagnosis technology and material source of marine fish iridescent virus, more specifically to a flounder lymphocyst virus virus At the same time, it also relates to a preparation method of a flounder lymphocyst virus strain and its use in the detection and diagnosis of fish lymphocyst virus and in the preparation of cell engineering vaccines of seawater fish iridescent virus. Background technique [0002] Flounder (Paralichthys olivaceus, Japanese flounder) is an economical marine fish that is popular with consumers and has a promising market prospect. However, with the intensification and rapid development of mariculture industry, lymphocystis disease of flounder, which is caused by Lymphocystis disease virus (LCDV), is spreading rapid...

Claims

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Application Information

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IPC IPC(8): A61K39/12C12N7/01C12Q1/70
Inventor 张奇亚李正秋袁秀平桂建平
Owner INST OF AQUATIC LIFE ACAD SINICA
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