Autor(es): Bruno John G, Carrillo Maria P, Richarte Alicia M, Phillips Taylor, Andrews Carrie, Lee John S

Resumo: Nucleic acid aptamers have long demonstrated the capacity to bind viral envelope proteins - to inhibit the progression of pathogenic virus infections. Here we report on initial efforts to develop - screen DNA aptamers against recombinant envelope proteins or synthetic peptides - whole inactivated viruses from several virulent arboviruses including Chikungunya, Crimean-Congo hemorrhagic fever (CCHF), dengue, tickborne encephalitis - West Nile viruses. We also analyzed sequence data - secondary structures for commonalities that might reveal consensus binding sites among the various aptamers. Some of the highest affinity - most specific aptamers in the down-selected libraries were demonstrated to have diagnostic utility in lateral flow chromatographic assays - in a fluorescent aptamer-magnetic bead s-wich assay. Some of the reported aptamers may also be able to bind viral envelope proteins in vivo - therefore may have antiviral potential in passive immunity or prophylactic applications. 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 - secondary structures of top-ranked anti-arboviral aptamers suggest potential virus binding motifs exist within some of the key aptamers - are highlighted in the suplemental 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 - only a few other times in a larger library of aptamers known to bind bacteria - rickettsia or other targets. Diagnostic utility of some of the aptamers for arbovirus detection in lateral flow chromatographic assays - a fluorescent s-wich assay on the surface of magnetic microbeads is also demonstrated. This article catalogues numerous DNA aptamer sequences which can bind various important pathogenic arboviruses - have, in some cases, already demonstrated diagnostic potential. These aptamer sequences are proprietary, patent-pending, - partially characterized. Therefore, they are offered to the scientific community for potential research use in diagnostic assays, biosensor applications or for possible passive immunity - prophylaxis against pathogenic viruses.

Palavras-Chave: Aptamer; Arbovirus; Consensus; DNA; Enzyme-linked; Fluorescence; Lateral flow; SELEX

Imprenta: BMC Research Notes, v. 5, p. 633, 2012

Identificador do objeto digital: 10.1186/1756-0500-5-633

Descritores: Chikungunya virus - Arbovirus ; Chikungunya virus - DNA ; Chikungunya virus - Molecular structure ; Chikungunya virus - Pathogenesis ; Chikungunya virus - Proteins ; Chikungunya virus - Viral infections ; Chikungunya virus - Molecular methods ; Chikungunya Virus - Virus ; Chikungunya virus - Molecular screening ; Chikungunya virus - Chikungunya fever ; Chikungunya virus - Dengue ; Chikungunya virus - Immunology

Data de publicação: 2012