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Journal of Biomedicine and Biotechnology
Volume 2010, Article ID 934694, 8 pages
http://dx.doi.org/10.1155/2010/934694
Research Article

Characterization of Recombinant Dengue-2 Virus Derived from a Single Nucleotide Substitution in the 5′ Noncoding Region

1Department of Anatomy, Faculty of Science, Mahidol University, Bangkok 10400, Thailand
2Arbovirus Diseases Branch, Division of Vector-Borne Infectious Diseases, Center for Disease Control and Prevention, Fort Collins, CO 80522, USA

Received 25 May 2009; Revised 15 October 2009; Accepted 23 December 2009

Academic Editor: Shahid Jameel

Copyright © 2010 Vijittra Leardkamolkarn et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Variants of wild-type dengue serotype 2 (DEN-2) virus containing nucleotide substitutions at positions 14, 15, or 57 in the NCR were constructed by PCR-mediated site-directed mutagenesis. All three viruses containing a single point substitution demonstrated attenuation phenotype as evidenced by decreases replication and plaque size in cell culture assay. All three variants were less neurovirulent in newborn mice compared to the wild type. The mutants were immunogenic in adult mice immunogenicity and maintained stable replication characteristics following passage in mice. The variant viruses were competent for replication in Aedes aegypi mosquito vector, albeit at lower levels of infection and dissemination in the mosquito than the wild-type Den-2 16681 virus. Although all of the viruses, including the wild type, were found transmissible in mosquito life cycles, they were found subsequentially decreased in efficiency of infection, transmission, and dissemination rates along the mosquito generations and all of them remained genetically stable.