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.

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.