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Development, screening, and analysis of DNA aptamer libraries potentially useful for diagnosis and passive immunity of arboviruses.
- Source :
-
BMC research notes [BMC Res Notes] 2012 Nov 13; Vol. 5, pp. 633. Date of Electronic Publication: 2012 Nov 13. - Publication Year :
- 2012
-
Abstract
- Background: Nucleic acid aptamers have long demonstrated the capacity to bind viral envelope proteins and to inhibit the progression of pathogenic virus infections. Here we report on initial efforts to develop and screen DNA aptamers against recombinant envelope proteins or synthetic peptides and whole inactivated viruses from several virulent arboviruses including Chikungunya, Crimean-Congo hemorrhagic fever (CCHF), dengue, tickborne encephalitis and West Nile viruses. We also analyzed sequence data and secondary structures for commonalities that might reveal consensus binding sites among the various aptamers. Some of the highest affinity and most specific aptamers in the down-selected libraries were demonstrated to have diagnostic utility in lateral flow chromatographic assays and in a fluorescent aptamer-magnetic bead sandwich assay. Some of the reported aptamers may also be able to bind viral envelope proteins in vivo and therefore may have antiviral potential in passive immunity or prophylactic applications.<br />Results: Several arbovirus DNA aptamer sequences emerged multiple times in the various down selected aptamer libraries thereby suggesting some consensus sequences for binding arbovirus envelope proteins. Screening of aptamers by enzyme-linked aptamer sorbent assay (ELASA) was useful for ranking relative aptamer affinities against their cognate viral targets. Additional study of the aptamer sequences and secondary structures of top-ranked anti-arboviral aptamers suggest potential virus binding motifs exist within some of the key aptamers and are highlighted in the supplemental figures for this article. One sequence segment (ACGGGTCCGGACA) emerged 60 times in the anti-CCHF aptamer library, but nowhere else in the anti-arbovirus library and only a few other times in a larger library of aptamers known to bind bacteria and rickettsia or other targets. Diagnostic utility of some of the aptamers for arbovirus detection in lateral flow chromatographic assays and a fluorescent sandwich assay on the surface of magnetic microbeads is also demonstrated.<br />Conclusions: This article catalogues numerous DNA aptamer sequences which can bind various important pathogenic arboviruses and have, in some cases, already demonstrated diagnostic potential. These aptamer sequences are proprietary, patent-pending, and partially characterized. Therefore, they are offered to the scientific community for potential research use in diagnostic assays, biosensor applications or for possible passive immunity and prophylaxis against pathogenic viruses.
- Subjects :
- Aptamers, Nucleotide isolation & purification
Arbovirus Infections diagnosis
Arbovirus Infections virology
Base Sequence
Chromatography methods
Enzyme-Linked Immunosorbent Assay
Gene Library
Humans
Immunization, Passive methods
Molecular Sequence Data
SELEX Aptamer Technique methods
Spectrophotometry
Viral Proteins immunology
Aptamers, Nucleotide genetics
Aptamers, Nucleotide immunology
Arbovirus Infections immunology
Arboviruses immunology
Subjects
Details
- Language :
- English
- ISSN :
- 1756-0500
- Volume :
- 5
- Database :
- MEDLINE
- Journal :
- BMC research notes
- Publication Type :
- Academic Journal
- Accession number :
- 23148669
- Full Text :
- https://doi.org/10.1186/1756-0500-5-633