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Method for preparing intestinal virus vaccine by using gerbil kidney cell

An enterovirus and kidney cell technology, applied in animal cells, antiviral agents, vertebrate cells, etc., can solve the problems of low reproductive titer, low efficiency, yield limitation, etc., and achieve the effect of convenient purification and simple production process.

Active Publication Date: 2014-07-16
ZHEJIANG CENT FOR DISEASE CONTROL & PREVENTION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For the former, the reproduction titer of EV71 virus in this diploid cell is low, the yield is limited, and it is difficult to expand production; for the latter, since Vero cells are passaged cells, the DNA content of passaged cells contained in the vaccine needs to be below 30ng. However, it is difficult to remove the DNA in the virus fluid, resulting in higher production costs and lower efficiency

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Embodiment 1: Preparation of EV71 monovalent vaccine

[0021] (1) Preparation of cell monolayer

[0022] Take gerbils aged 10-15 days, cut off the carotid artery and bleed to death, soak in 0.2% (v / v) new clean and disinfectant solution for 30 minutes, take the kidney aseptically, cut it into pieces, and contain 100U / ml kanamycin Wash three times with MEM culture solution, centrifuge to remove the washing solution; add 0.25% (w / v) trypsin digestion solution 10 times the weight of the kidney, and digest at 4°C; discard the digestion solution after 16 hours, and wash once with MEM culture solution . Add MEM solution 10 times the weight of the kidney to disperse the cells by pipetting repeatedly, add newborn bovine serum to a final concentration of 3% (v / v), inoculate the cell bottle, and culture at 36°C for 2-3 days. When the cell growth reached 50%-60% confluence, the MEM culture with 3% (v / v) newborn bovine serum was replaced to continue the culture. When the cells r...

Embodiment 2

[0034] Embodiment 2: Preparation of CoxA16 vaccine

[0035] In Example 1, the inoculated virus was replaced with the CoxA16 virus strain (provided by Hangzhou Sixth People's Hospital), and the other preparation methods were the same, and CoxA16 vaccine semi-finished products and finished vaccines could be prepared.

Embodiment 3

[0036] Embodiment 3: Preparation of EV71 and CoxA16 bivalent vaccine

[0037] The EV71 vaccine semi-finished product prepared in Example 1 and the CoxA16 vaccine semi-finished product prepared in Experimental Example 2 were mixed at a volume ratio of 1:1 and subpackaged to obtain a bivalent enterovirus vaccine.

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PUM

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Abstract

The invention provides a method for preparing an intestinal virus vaccine by using a gerbil kidney cell as a matrix cell for virus culture. According to the method, the gerbil kidney cell is used as the matrix cell for culturing EV71 or CoxA16 virus, wherein the growth titer of the EV71 or CoxA16 virus on the cell is high and liquid can be collected for many times. Secondly, the cell is a primary cell, so that the problem of difficulty in treatment of DNA (Deoxyribonucleic Acid) of a passage cell is solved; a production process for preparing the vaccine is relatively simple; the purification is convenient; and the prepared vaccine is safer and more reliable for inoculation of infants and young children.

Description

(1) Technical field [0001] The invention relates to a method for preparing enterovirus vaccine using gerbil kidney cells. (2) Background technology [0002] HFMD can be caused by a variety of enteroviruses, including CoxA5, A10, A16, A19, EV71, and some echoviruses and Coxsackie group B viruses, among which CoxA16 and EV71 are the most common. [0003] Enterovirus 71 (EV71) belongs to the genus Human Enterovirus, Picornaviridae, and is one of the main pathogens of hand, foot and mouth disease. Hand, foot, mouth and herpetic angina after infection in infants and young children, a small number of complications such as aseptic meningitis, brainstem encephalitis, neurogenic pulmonary edema and acute flaccid paralysis, severe nervous system, respiratory system and circulatory symptoms. Since the virus was first isolated in California, USA in 1969, the world has experienced many outbreaks of EV71. Since 1997, the number of people infected with EV71 has increased significantly, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/125A61K39/295A61P31/14C12N5/071
Inventor 朱函坪朱智勇蒋健敏钱磊徐芳姚苹苹杨章女谢荣辉周小龙
Owner ZHEJIANG CENT FOR DISEASE CONTROL & PREVENTION
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