KMB17 cell-adapted strain for II type dengue virus and preparation method thereof

A technology of dengue virus and adapted strain, applied in the field of virus vaccine, to achieve the effect of good antigenicity, huge economic value and social benefit

Inactive Publication Date: 2013-02-06
INST OF MEDICAL BIOLOGY CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Human embryonic lung diploid cell KMB17 has been successfully used in the production of hepatitis A vaccine and polio vaccine, and its safety and cell stability have been widely verified, but it has not been successfully applied to the Chinese strain of dengue virus type Ⅱ. report

Method used

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  • KMB17 cell-adapted strain for II type dengue virus and preparation method thereof
  • KMB17 cell-adapted strain for II type dengue virus and preparation method thereof
  • KMB17 cell-adapted strain for II type dengue virus and preparation method thereof

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Embodiment 1

[0032]1) Aedes albopictus cells C6 / 36 were subcultured as follows: Add the following cell growth medium to the cells as usual: RPMI-1640 medium plus 10% volume ratio (v / v) calf serum; 28 + 2°C, in an environment with a carbon dioxide volume concentration of 5% (v / v), culture at a constant temperature, change the medium once every 2-3 days, and grow cells into a dense monolayer with good shape after 4-5 days;

[0033] 2) According to the amount of 1ml virus liquid / bottle, inoculate the type II dengue virus liquid with a concentration of 4.0mol on the densely grown and transparent Aedes albopictus cells C6 / 36 in step 1), and the cells in each bottle The quantity is 1~1.7×10 6 , at a temperature of 28 + 2°C, CO 2 In an environment with a volume concentration of 5% (v / v), adsorb at a constant temperature for 1 hour. In the same environment, add the following virus maintenance solution at a volume ratio of 1:9: RPMI-1640 medium + 2% volume ratio ( v / v) calf serum + 2% volume ra...

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Abstract

The invention discloses a KMB17 cell-adapted strain for the II type dengue virus and a preparation method thereof. The epidemic II type dengue virus strain can be adapted to the human embryo lung diploid cell KMB17 to obtain the cell-adapted strain with stable passage and high virus amplification capacity, and purified strains with strong virulence and purity are obtained through plague purification; after identification, the virus purified strains maintain basic biological characteristics of original strains, are provided with good antigenicity, and are capable of performing II type dengue virus amplification with the human embryo lung diploid cell KMB17 as the matrix, limitation of host cells of the II type dengue virus is broken, a new thinking with China's regional characteristics is opened up for research of the vaccine for preventing the dengue virus, the function is laid for research of inactivation of the II type dengue virus and attenuated live vaccines, feasibility of mass production is provided simultaneously, and the research and development of the vaccine can produce great economic value and social benefits.

Description

[0001] technical field [0002] The invention relates to a method for preparing a virus strain, in particular to a method for isolating, screening, breeding, purifying and cultivating an adapted strain of type II dengue virus on KMB17 cells, and belongs to the technical field of virus vaccines. Background technique [0003] Dengue virus is an arbovirus transmitted by Aedes aegypti and Aedes albopictus. It is widely prevalent in more than 100 tropical and subtropical countries and regions in the world. It can cause dengue fever, dengue hemorrhagic fever and more serious dengue Hemorrhagic shock syndrome. According to different antigenicity, dengue virus can be divided into four serotypes: I, II, III, and IV, among which type II is the most widely spread. In recent years, due to factors such as global warming and frequent population movements, the incidence of dengue fever in the world has increased by nearly 30 times. In the past two years, the number of people infected wit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00C12R1/93
Inventor 孙强明赵玉娇潘玥陈俊英杨丽娟岳耀斐黄新伟施海晶丁晓洁
Owner INST OF MEDICAL BIOLOGY CHINESE ACAD OF MEDICAL SCI
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