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High-growth enterovirus 71 strain and its vaccine

A technology of enterovirus and vaccine, applied in the direction of virus, virus/bacteriophage, virus antigen components, etc.

Active Publication Date: 2021-12-24
台湾卫生研究院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] Four of the five candidate vaccines use Vero cells (a cell line commonly used in the production of human vaccines), but the peak virus titer can only reach around 107TCID50 / ml (Chou et al.2012; Liang et al.2012)

Method used

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  • High-growth enterovirus 71 strain and its vaccine
  • High-growth enterovirus 71 strain and its vaccine
  • High-growth enterovirus 71 strain and its vaccine

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Experimental program
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experiment example

[0023] Domestication of High-Growing Vaccine Viruses in Vero Cells

[0024] method

[0025] Cells, media and virus: Enterovirus 71 was cultured in human rhabdomyosarcoma (RD) cells and African green monkey kidney (Vero) cells according to standard procedures (Wu et al., 2001, Huang et al., 2010).

[0026] Virology determination: virus infectivity titer test is 50% tissue culture infectious dose (TCID 50 ) to observe cytopathic effect (CPE), and measure plaque forming units (PFU) for plaque analysis (Huang et al., 2010). Also includes a positive control group with pre-set tolerance ranges for TCID 50 and plaque analysis.

[0027] Determination of the growth curve of the virus in cell culture: the EV71 virus was cultured in a 6-well culture dish, and the supernatant was obtained from the first to the sixth day after infection to determine the virus infection titer. A microcarrier cell culture system was performed in serum-containing media to evaluate the effect of using Vero...

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Abstract

The present invention relates to a domesticated enterovirus 71 (EV71) vaccine strain for improving vaccine production in mammals. The EV71 vaccine strain contains at least one modified enterovirus protein, and can rapidly propagate EV71 virus from mammalian host cells (such as Vero cells). The present invention also relates to a vaccine produced from a domesticated EV71 virus vaccine strain and a method for producing the vaccine.

Description

technical field [0001] The present invention relates to an isolated enterovirus type 71 (EV71) vaccine strain, in particular to an enterovirus type 71 (EV71) B5 genotype vaccine virus strain, which can be expressed in mammalian host cells (such as Vero cells) ) rapid growth, and induce cross-reactive neutralizing antibody titers against other genotypes of EV71 virus. Background technique [0002] Enterovirus type 71 (EV71), along with other important human pathogens such as polioviruses and rhinoviruses, is a positive-sense RNA virus belonging to the family Picornaviridae and the genus Enterovirus. EV71 was first isolated in 1969 from an American child with neurological disease, but according to a recent literature review, EV71 appeared in the Netherlands as early as 1963 (VanDer Saden et al., 2009). The EV71 genome is translated into a long polyprotein with a single open reading frame and a polyadenine sequence stretch (poly A tract), and there are untranslated regions at ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/12A61K39/00C12N7/04C07H21/04
CPCA61K39/12A61K2039/55505C12N2770/32321C12N2770/32334C12N7/00C12N2770/32322
Inventor 李敏西
Owner 台湾卫生研究院