EV-71 virus seed, inactivated vaccine for human and method of producing the same

An inactivated vaccine, EV-71 technology, applied in biochemical equipment and methods, antiviral agents, viruses/phages, etc., can solve the problems of non-specific vaccine application, achieve good growth and replication ability, and effective immunogenicity Effect

Active Publication Date: 2009-04-08
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

But so far, although the virus infection has caused more morbidity in various regions of my country, there is still no effective specific vaccine application.

Method used

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  • EV-71 virus seed, inactivated vaccine for human and method of producing the same
  • EV-71 virus seed, inactivated vaccine for human and method of producing the same
  • EV-71 virus seed, inactivated vaccine for human and method of producing the same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] ——Collection and identification of EV-71 inactivated vaccine virus FY-23 virus

[0039] The secretions taken from the cotton swabs of the respiratory tract of critically ill patients with HFMD were inoculated in Vero cells, and then the same cells were subcultured twice, and then replanted with KMB-17 cells to grow to the seventh generation. Take 1ml of the virus liquid, perform RT-PCR according to the conventional method, obtain the cDNA sequence of the viral gene in segments, and connect the fragment into the puc-18 vector according to the conventional method, perform conventional nucleotide sequencing analysis, and use this sequence in the The results were confirmed by comparing with similar gene pools. Secondly, use anti-EV-71 specific serum, after diluting 1:4, mix it with 10-fold dilution of the virus in equal amounts, place it at 37°C for 1 hour, then inoculate Vero cells or KMB17 cells, and observe the lesion after culturing at 37°C for one week As a result, an...

Embodiment 2

[0041] The KMB-17 cell line was inoculated with the suspension adsorption method of the 15th generation working seed of the EV-71 vaccine virus, and cultured and grown at 37°C for 5 days. Analysis and purification, content determination, relevant dilution ratio, the vaccine prepared by adding aluminum hydroxide adsorption, consists of the following components: EV-71 inactivated purified antigen 100μg / ml, aluminum hydroxide 0.8-1mg / ml, thimerosal 0.05-0.1ng / ml.

Embodiment 3

[0043] ——Suspension adsorption method in the preparation process of EV-71 inactivated vaccine

[0044] Take densely growing single-layer KMB-17 cells, digest them with 0.1% trypsin, remove the digestive juice, suspend the cells at 5% of the volume of the original culture medium, add 0.2-0.5ml virus solution, the moi is 0.05, mix well Incubate at 37°C for 30 minutes and shake gently in it, then transfer the suspension into a culture bottle, add an appropriate amount of growth medium, and culture at 37°C until the cells adhere to the wall and the lesions are completely harvested.

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Abstract

The invention discloses an EV-71 virus species, a human inactivated vaccine and a preparation method thereof. The classification name of the virus species is intestinal virus 71 with a storage number of CGMCC No.2701. The prepared human EV-71 inactivated vaccine comprises the following components: 100 Mu g / ml of EV-71 alive purified antigen, 0.8-1mg / ml of aluminum hydroxide, and 0.05-0.1mg / ml of merthiolate. The experiments show that the vaccine products have good immunogenicity and safety.

Description

technical field [0001] The invention belongs to the technical field of human preventive medicine. More specifically, the present invention relates to virus seeds and methods for their preparation. Simultaneously, the present invention also relates to an inactivated vaccine for humans made from the virus seeds and a preparation method thereof. Background technique [0002] EV-71 (Enterovirus 71) infection is one of the main forms of HFMD. EV-71 belongs to the enterovirus family and is a small RNA virus with a typical icosahedral structure and no envelope. It is usually transmitted through intestinal excretion→oral, and under certain circumstances, it can also be transmitted through the respiratory tract. Infection caused by this virus usually manifests as characteristic herpes on the hands, feet and mouth, accompanied by a certain degree of fever and cold symptoms. In severe cases, it can cause lesions in the nervous system and respiratory system, and has a certain mortali...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00C12N7/06A61K39/125A61P31/14
Inventor 李琦涵董承红刘龙丁谢忠平王丽春马绍辉崔萍芳梁燕刘云霞唐松清
Owner INST OF MEDICAL BIOLOGY CHINESE ACAD OF MEDICAL SCI
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