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- W4313373622 abstract "Abstract Objective Reverse genetic system have been used to generate mutant viruses to investigate the genetic features as well as to produce vaccine strain of various viruses. However, the virus producing system has not been applied to produce vaccine strain of Hepatitis A virus (HAV). Here, we established reverse genetic system to generate vaccine strain of HAV and assessed the generated HAV in terms of immunogenicity. Methods To produce HAV, MRC-5 cells were transfected with HAV RNA generated via in vitro transcription. To confirm the virus generation, we performed RT-PCR, Western blot, Rapid amplification of cDNA ends (RACE) and transmission electron microscopy (TEM) analysis using the supernatant of the HAV RNA transfected cells. To assess the immunogenicity of the generated HAV, mice were immunized with the generated and inactivated HAV and performed enzyme-linked immunosorbent assay (ELISA) to assess anti-HAV IgG using sera from the immunized mice. Results From the supernatant of HAV RNA transfected cells, HAV-specific RNA and protein were detected. TEM analysis showed that there are two types of HAV particles as shown in a previous report: naked and enveloped HAV, which is one of HAV features. Although seven nucleotide sequences are different in the coding sequence for structural protein compared to those of wildtype vaccine strain of HAV, genomic RNA analysis using RACE showed that the generated HAV have HAV-specific RNA as genome. In the immunogenicity study using mouse, the generated HAV induced HAV-specific IgG and the titer was comparable to wild-type vaccine strain of HAV. Conclusions We established the reverse genetic system to produce vaccine strain of HAV and confirmed that the generated HAV can be used as vaccine strain." @default.
- W4313373622 created "2023-01-06" @default.
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- W4313373622 date "2020-05-01" @default.
- W4313373622 modified "2023-09-27" @default.
- W4313373622 title "Development of a reverse genetics system to generate vaccine strain of Hepatitis A virus" @default.
- W4313373622 doi "https://doi.org/10.4049/jimmunol.204.supp.166.25" @default.
- W4313373622 hasPublicationYear "2020" @default.
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