Dengue Fever Research Today is a free monthly online journal that collates and summarizes the latest research about Dengue Fever, including details on symptoms, risk factors, diagnosis, treatment, prevention. | ||||||||
|
Suckling mice were used to detect infectious dengue-2 viruses by intracerebral injection of the full-length RNA transcript.Lee YR, Huang KJ, Lei HY, Chen SH, Lin YS, Yeh TM, Liu HS Department of Microbiology and Immunology, College of Medicine, National Cheng Kung University, Tainan, Taiwan. 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. Published 6 April 2005 in Intervirology, 48(2): 161-6.
© 2004-2008 Dengue Fever Research Today. All Rights Reserved. |
| ||||||
