Suckling Mice Were Used to Detect Infectious Dengue-2 Viruses by Intracerebral Injection of the Full-Length RNA Transcript

Vol. 48, No. 2-3, 2005

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Original Paper

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

Department of Microbiology and Immunology, College of Medicine, National Cheng Kung University, Tainan, Taiwan

Address of Corresponding Author

Intervirology 2005;48:161-166 (DOI: 10.1159/000081744)

Key Words

Dengue-2-virus
Full-length cDNA
Suckling mice system

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 vitrotranscription was usedto synthesizefull-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: Wedeveloped 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.

Author Contacts

H.-S. Liu
Department of Microbiology and Immunology
College of Medicine, National Cheng Kung University
1 Da-Shue Road, Tainan 701 (Taiwan)
Tel. +886 6 235 3535, ext. 5630, Fax +886 6 208 2705, E-Mail a713@mail.ncku.edu.tw

Article Information

Received: January 22, 2004
Accepted after revision: July 5, 2004
Number of Print Pages : 6
Number of Figures : 4, Number of Tables : 1, Number of References : 23

Medline Abstract (ID 15812190)

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