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Characterization of DNA aptamers generated against the soft-shelled turtle iridovirus with antiviral effects

BACKGROUND: Soft-shelled turtle iridovirus (STIV) causes severe systemic disease in farmed soft-shelled turtles (Trionyx sinensis). More efficient methods of controlling and detecting STIV infections are urgently needed.  METHODS: In this study, we generated eight single-stranded DNA (ssDNA) aptamer...

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Autores principales: Li, Pengfei, Zhou, Lingli, Yu, Yepin, Yang, Min, Ni, Songwei, Wei, Shina, Qin, Qiwei
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4588899/
https://www.ncbi.nlm.nih.gov/pubmed/26419355
http://dx.doi.org/10.1186/s12917-015-0559-6
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author Li, Pengfei
Zhou, Lingli
Yu, Yepin
Yang, Min
Ni, Songwei
Wei, Shina
Qin, Qiwei
author_facet Li, Pengfei
Zhou, Lingli
Yu, Yepin
Yang, Min
Ni, Songwei
Wei, Shina
Qin, Qiwei
author_sort Li, Pengfei
collection PubMed
description BACKGROUND: Soft-shelled turtle iridovirus (STIV) causes severe systemic disease in farmed soft-shelled turtles (Trionyx sinensis). More efficient methods of controlling and detecting STIV infections are urgently needed.  METHODS: In this study, we generated eight single-stranded DNA (ssDNA) aptamers against STIV using systematic evolution of ligands by exponential enrichment (SELEX). RESULTS: The aptamers formed representative stem-loop secondary structures. Electrophoretic mobility shift assays and fluorescent localization showed that the selected aptamers had high binding affinity for STIV. Aptamer QA-36 had the highest calculated binding affinity (K(d)) of 53.8 nM. Flow cytometry and fluorescence microscopy of cell-aptamer interactions demonstrated that QA-12 was able to recognize both STIV-infected cells and tissues with a high level of specificity. Moreover, the selected aptamers inhibited STIV infection in vitro and in vivo, with aptamer QA-36 demonstrating the greatest protective effect against STIV and inhibiting STIV infection in a dose-dependent manner. DISCUSSION: We generated DNA aptamers that bound STIV with a high level of specificity, providing an alternative means for investigating STIV pathogenesis, drug development, and medical therapies for STIV infection. CONCLUSIONS: These DNA aptamers may thus be suitable antiviral candidates for the control of STIV infections.
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spelling pubmed-45888992015-10-01 Characterization of DNA aptamers generated against the soft-shelled turtle iridovirus with antiviral effects Li, Pengfei Zhou, Lingli Yu, Yepin Yang, Min Ni, Songwei Wei, Shina Qin, Qiwei BMC Vet Res Research Article BACKGROUND: Soft-shelled turtle iridovirus (STIV) causes severe systemic disease in farmed soft-shelled turtles (Trionyx sinensis). More efficient methods of controlling and detecting STIV infections are urgently needed.  METHODS: In this study, we generated eight single-stranded DNA (ssDNA) aptamers against STIV using systematic evolution of ligands by exponential enrichment (SELEX). RESULTS: The aptamers formed representative stem-loop secondary structures. Electrophoretic mobility shift assays and fluorescent localization showed that the selected aptamers had high binding affinity for STIV. Aptamer QA-36 had the highest calculated binding affinity (K(d)) of 53.8 nM. Flow cytometry and fluorescence microscopy of cell-aptamer interactions demonstrated that QA-12 was able to recognize both STIV-infected cells and tissues with a high level of specificity. Moreover, the selected aptamers inhibited STIV infection in vitro and in vivo, with aptamer QA-36 demonstrating the greatest protective effect against STIV and inhibiting STIV infection in a dose-dependent manner. DISCUSSION: We generated DNA aptamers that bound STIV with a high level of specificity, providing an alternative means for investigating STIV pathogenesis, drug development, and medical therapies for STIV infection. CONCLUSIONS: These DNA aptamers may thus be suitable antiviral candidates for the control of STIV infections. BioMed Central 2015-09-30 /pmc/articles/PMC4588899/ /pubmed/26419355 http://dx.doi.org/10.1186/s12917-015-0559-6 Text en © Li et al. 2015 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
Li, Pengfei
Zhou, Lingli
Yu, Yepin
Yang, Min
Ni, Songwei
Wei, Shina
Qin, Qiwei
Characterization of DNA aptamers generated against the soft-shelled turtle iridovirus with antiviral effects
title Characterization of DNA aptamers generated against the soft-shelled turtle iridovirus with antiviral effects
title_full Characterization of DNA aptamers generated against the soft-shelled turtle iridovirus with antiviral effects
title_fullStr Characterization of DNA aptamers generated against the soft-shelled turtle iridovirus with antiviral effects
title_full_unstemmed Characterization of DNA aptamers generated against the soft-shelled turtle iridovirus with antiviral effects
title_short Characterization of DNA aptamers generated against the soft-shelled turtle iridovirus with antiviral effects
title_sort characterization of dna aptamers generated against the soft-shelled turtle iridovirus with antiviral effects
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4588899/
https://www.ncbi.nlm.nih.gov/pubmed/26419355
http://dx.doi.org/10.1186/s12917-015-0559-6
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