Rabies virus CTN chick-embryo cell adaptive strain

A technology of rabies virus and chicken embryo cells, applied in the field of new rabies virus strains, to achieve good immunogenicity, weakened pathogenicity, and safe and reliable effects

Active Publication Date: 2014-06-18
SHENZHEN WEIGUANG BIOLOGICAL PROD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

To date, there is no literature mentioning the adaptation of CTN-1 strains for CEC

Method used

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  • Rabies virus CTN chick-embryo cell adaptive strain
  • Rabies virus CTN chick-embryo cell adaptive strain
  • Rabies virus CTN chick-embryo cell adaptive strain

Examples

Experimental program
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Effect test

preparation example Construction

[0018] The preparation of CEC, the Fluorescence Focus Units Assay (FFU) and the preparation of phosphate buffered saline (PBS) are involved several times below, and the specific operations are as follows:

[0019] (1) Preparation of primary chicken embryo fibroblasts (CECs)

[0020] Select eggs that have been produced within one week, have normal shape, uniform eggshell thickness, no cracks, and thick egg whites, and put them in an incubator at 38±1°C and relative humidity of 60±20% for incubation, and observe with an egg inspection lamp Whether it is a fertilized egg and its vitality. Choose 9-11 days old chicken embryos with normal development, visible blood vessels and activities. Soak it in 0.2% (m / v) bromine solution for 5 minutes, remove it, put the air cell up on the egg tray, and then disinfect it with 2% iodine (m / v) and 75% alcohol (v / v) Inside the clean bench. Take out the chicken embryo with sterile tweezers and put it into a plate filled with 1× Hanks solution....

Embodiment 1

[0038] Taking the CTN-1V5 strain as the mother strain, through the comparison and screening of multiple passage ways and processes, it was found that the strains obtained by passage according to the following process routes can gradually adapt to the growth in chicken embryo cells. The chicken embryo cell-adapted strain of rabies virus It was named CTNCEC25 strain.

[0039] Step 1: subculture the CTN-1V5 strain on vero cells to increase the virus titer.

[0040] Use PBS (pH7.4) to make 10-fold serial dilutions of RV CTN-1V5 (from the China Institute for Food and Drug Control, the 5th generation virus of CTN-1 strain vero cells), and then follow the ratio of 1:100 to 1:1000. Proportionally inoculate vero monolayer cells (from China National Institutes for Food and Drug Control, passage 121), absorb at 37°C for 60 minutes, and then add cell maintenance solution (based on 199 medium, add 10% (v / v) of final concentration Bovine serum, pH7.2~8.0), placed at 37℃, 5%CO 2 Cultivate ...

Embodiment 2

[0048] The relevant preservation information of the RV CTNCEC25 strain of the present invention is as follows:

[0049] Classification name: rabies virus (Rabies virus).

[0050] Deposit date: September 14, 2012.

[0051] Name of depository unit: General Microbiology Center of China Committee for the Collection of Microorganisms.

[0052] Abbreviation of depository unit: CGMCC.

[0053] Address of depository unit: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, China, zip code 100101.

[0054] Deposit number: 6510.

[0055] Taxonomy: A new strain belonging to Rhabdoviridae, genus Lyssaviruses.

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Abstract

The invention provides a rabies virus CTN chick-embryo cell adaptive strain that the microbial preservation number is CGMCC No.6510. The invention further provides an application of the rabies virus CTN chick-embryo cell adaptive strain in preparing rabies virus inactivated vaccine. The rabies virus CTN chick-embryo cell adaptive strain has a novel alkali radical and an amino acid sequence, can generate high virus titer on CEC (Cation Exchange Capacity), has high immunogenicity, can be used for producing rabies vaccine, preparing a RV (Rota Virus) antibody, detecting the valence of the RV antibody and the like. Five structural protein and complete genome sequences of the strain are detected. Besides, the invention further provides inactivation superantigen vaccine of a RV CTNCEC25 strain. The strain has good immunizing protection, the potency can meet the level of vaccine in market at present, and the pharmacopeia standards are met.

Description

technical field [0001] The invention mainly relates to a novel rabies virus strain, in particular to a chicken embryo cell-adapted strain of the rabies virus. Background technique [0002] Rabies virus (RV) is a highly neurotropic virus. It is a non-segmented single-stranded negative-sense RNA virus in the Rhabdoviridae (Lyssavirus genus), which can cause zoonosis in the world Sexually transmitted diseases - rabies. It is reported that there are about 55,000 deaths due to rabies every year in the world, and the actual number of deaths should be significantly higher than the statistics (http: / / www.worldrabiesday.org / ). At present, except for Japan, the United Kingdom, Hawaii and a few other countries and regions where no rabies occurs, the disease is prevalent worldwide. Among them, Asia and Africa are the regions where human rabies occurs most seriously, accounting for 99% of the total number of deaths in the world. Among countries that officially report rabies, India rank...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A61K39/205A61P31/14C12R1/93
CPCA61K39/12A61K2039/5252A61P31/14C12N1/00C12N7/00C12N2760/20121C12N2760/20134C12R2001/91
Inventor 王春华郭采平罗姗刘永娣李慧丁玉江容伟华周兰贞周维宋清爽黄伟荣田华朱士茂张信
Owner SHENZHEN WEIGUANG BIOLOGICAL PROD
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