Suckling mice were used to detect infectious dengue-2 viruses by intracerebral injection of the full-length RNA transcript.

Abstract:

OBJECTIVE:In this study we established a mouse brain injection system to detect infectious dengue-2 virus produced from the full-length RNA transcripts. METHODS:In vitro transcription was used to synthesize full-length dengue-2 virus RNA from the plasmid pRS424FLDEN2NGC, which was intracerebrally injected into the 6-day-old suckling mice (ICR strains). Engineered dengue-2 viruses were detected in the brain sections using immunohistochemistry staining. RT-PCR followed by restriction endonuclease BstEII digestion was used to confirm the mosquito C6/36 cells cocultured with the mouse brain extract. RESULTS:The mice inoculated with the full-length dengue-2 viral RNA transcript showed paralysis symptoms and died between day 10 and 13 postinjection. The dengue-2 virus-specific antigens (E, Core and NS1) were detected in all the brain and part of the liver sections of the paralyzed mice by immunohistochemistry staining, indicating the existence of dengue-2 virus in these tissues of the suckling mice. The viruses detected in the brains of suckling mice were indeed infectious, which was further confirmed by coculturing mosquito C6/36 cells with the brain extract of the injected mice. CONCLUSIONS:We developed an in vivo approach to detect and produce engineered dengue viruses with infectivity from the full-length plasmid cDNA. This suckling mice system will also aid in screening the infectious viruses that are created by site-directed mutagenesis and is useful for the studies of dengue virus gene function and pathogenesis in the host.

journal_name

Intervirology

journal_title

Intervirology

authors

Lee YR,Huang KJ,Lei HY,Chen SH,Lin YS,Yeh TM,Liu HS

doi

10.1159/000081744

keywords:

subject

Has Abstract

pub_date

2005-01-01 00:00:00

pages

161-6

issue

2-3

eissn

0300-5526

issn

1423-0100

pii

81744

journal_volume

48

pub_type

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