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

Ying Ray Lee, Kao Jean Huang, Huan Yao Lei, Shun Hua Chen, Yee Shin Lin, Trai Ming Yeh, Hsiao Sheng Liu

Research output: Contribution to journalArticlepeer-review

20 Citations (Scopus)

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 BstEll 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.

Original languageEnglish
Pages (from-to)161-166
Number of pages6
JournalIntervirology
Volume48
Issue number2-3
DOIs
Publication statusPublished - 2005 Mar 1

All Science Journal Classification (ASJC) codes

  • Virology
  • Infectious Diseases

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