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

Variants of wild-type dengue serotype 2 (DEN-2) virus containing nucleotide substitutions at positions 14, 15, or 57 in the 5′ NCR were constructed by PCR-mediated site-directed mutagenesis. All three viruses containing a single point substitution demonstrated attenuation phenotype as evidenced by d...

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Autores principales: Leardkamolkarn, Vijittra, Sirigulpanit, Wipawan, Kinney, Richard M.
Formato: Texto
Lenguaje:English
Publicado: Hindawi Publishing Corporation 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2843905/
https://www.ncbi.nlm.nih.gov/pubmed/20339476
http://dx.doi.org/10.1155/2010/934694
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author Leardkamolkarn, Vijittra
Sirigulpanit, Wipawan
Kinney, Richard M.
author_facet Leardkamolkarn, Vijittra
Sirigulpanit, Wipawan
Kinney, Richard M.
author_sort Leardkamolkarn, Vijittra
collection PubMed
description Variants of wild-type dengue serotype 2 (DEN-2) virus containing nucleotide substitutions at positions 14, 15, or 57 in the 5′ 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.
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spelling pubmed-28439052010-03-25 Characterization of Recombinant Dengue-2 Virus Derived from a Single Nucleotide Substitution in the 5′ Noncoding Region Leardkamolkarn, Vijittra Sirigulpanit, Wipawan Kinney, Richard M. J Biomed Biotechnol Research Article Variants of wild-type dengue serotype 2 (DEN-2) virus containing nucleotide substitutions at positions 14, 15, or 57 in the 5′ 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. Hindawi Publishing Corporation 2010 2010-03-22 /pmc/articles/PMC2843905/ /pubmed/20339476 http://dx.doi.org/10.1155/2010/934694 Text en 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.
spellingShingle Research Article
Leardkamolkarn, Vijittra
Sirigulpanit, Wipawan
Kinney, Richard M.
Characterization of Recombinant Dengue-2 Virus Derived from a Single Nucleotide Substitution in the 5′ Noncoding Region
title Characterization of Recombinant Dengue-2 Virus Derived from a Single Nucleotide Substitution in the 5′ Noncoding Region
title_full Characterization of Recombinant Dengue-2 Virus Derived from a Single Nucleotide Substitution in the 5′ Noncoding Region
title_fullStr Characterization of Recombinant Dengue-2 Virus Derived from a Single Nucleotide Substitution in the 5′ Noncoding Region
title_full_unstemmed Characterization of Recombinant Dengue-2 Virus Derived from a Single Nucleotide Substitution in the 5′ Noncoding Region
title_short Characterization of Recombinant Dengue-2 Virus Derived from a Single Nucleotide Substitution in the 5′ Noncoding Region
title_sort characterization of recombinant dengue-2 virus derived from a single nucleotide substitution in the 5′ noncoding region
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2843905/
https://www.ncbi.nlm.nih.gov/pubmed/20339476
http://dx.doi.org/10.1155/2010/934694
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