title: Selection of DNA aptamers that bind to influenza A viruses with high affinity and broad subtype specificity.
abstract: Many cases of influenza are reported worldwide every year. The influenza virus often acquires new antigenicity, which is known as antigenic shift; this results in the emergence of new virus strains, for which preexisting immunity is not found in the population resulting in influenza pandemics. In the event a new strain emerges, diagnostic tools must be developed rapidly to detect the novel influenza strain. The generation of high affinity antibodies is costly and takes time; therefore, an alternative detection system, aptamer detection, provides a viable alternative to antibodies as a diagnostic tool. In this study, we developed DNA aptamers that bind to HA1 proteins of multiple influenza A virus subtypes by the SELEX procedure. To evaluate the binding properties of these aptamers using colorimetric methods, we developed a novel aptamer-based sandwich detection method employing our newly identified aptamers. This novel sandwich enzyme-linked aptamer assay successfully detected the H5N1, H1N1, and H3N2 subtypes of influenza A virus with almost equal sensitivities. These findings suggest that our aptamers are attractive candidates for use as simple and sensitive diagnostic tools that need sandwich system for detecting the influenza A virus with broad subtype specificities.
authors.affiliation: VALWAY Technology Center, NEC Soft, Ltd., 1-18-7, Shinkiba Koto-ku, Tokyo 136-8627, Japan. Electronic address: shiratori-ikuo@mxh.nes.nec.co.jp.
authors.forename: Ikuo
authors.initials: I
authors.lastname: Shiratori
authors.affiliation: VALWAY Technology Center, NEC Soft, Ltd., 1-18-7, Shinkiba Koto-ku, Tokyo 136-8627, Japan.
authors.forename: Joe
authors.initials: J
authors.lastname: Akitomi
authors.affiliation: Division of Microbiology and Molecular Biology, IIT Research Institute, Illinois Institute of Technology, 10W. 35th Street, Chicago, IL 60616, United States.
authors.forename: David A
authors.initials: DA
authors.lastname: Boltz
authors.affiliation: VALWAY Technology Center, NEC Soft, Ltd., 1-18-7, Shinkiba Koto-ku, Tokyo 136-8627, Japan.
authors.forename: Katsunori
authors.initials: K
authors.lastname: Horii
authors.affiliation: VALWAY Technology Center, NEC Soft, Ltd., 1-18-7, Shinkiba Koto-ku, Tokyo 136-8627, Japan.
authors.forename: Makio
authors.initials: M
authors.lastname: Furuichi
authors.affiliation: VALWAY Technology Center, NEC Soft, Ltd., 1-18-7, Shinkiba Koto-ku, Tokyo 136-8627, Japan.
authors.forename: Iwao
authors.initials: I
authors.lastname: Waga
mesh_terms: D052157:Aptamers, Nucleotide / Q000737:chemistry / Q000302:isolation & purification*; D001483:Base Sequence; D004247:DNA / Q000737:chemistry*; D006801:Humans; D009980:Influenza A virus / Q000737:chemistry* / Q000302:isolation & purification; D007251:Influenza, Human / Q000175:diagnosis*; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D052156:SELEX Aptamer Technique; D020349:Surface Plasmon Resonance
chemical_list: D052157:Aptamers, Nucleotide; D004247:DNA
keywords: AIV; DNA aptamer; ELAA; Enzyme-linked aptamer assay (ELAA); HA; HEK; Influenza virus; MDCK; Madin-Darby canine kidney; NA; SELEX; SPR; avian influenza virus; enzyme-linked aptamer assay; hemagglutinin; human embryonic kidney; neuraminidase; surface plasmon resonance; systematic evolution of ligands by exponential enrichment