Selection and identification of an ssDNA aptamer against influenza B virus hemagglutinin protein

The influenza virus causes infectious respiratory disease with high morbidity and mortality worldwide. Influenza B typically goes unnoticed owing to its mild clinical symptoms and limitations. However, its increasing prevalence in recent years poses a significant health burden. Consequently, current diagnostic methods for the detection of influenza B virus are inadequate, highlighting the urgent need to develop accurate and sensitive techniques for early disease diagnosis. Aptamers, single-stranded deoxyribonucleic acid (ssDNA), or ribonucleic acid molecules primarily rely on their secondary structures, such as stem-loops and hairpins, to bind efficiently and specifically to the target through base complementary pairing, electrostatic interaction, hydrogen bonding, and van der Waals forces. Aptamers are superior to antibodies in their ability to bind targets. The objective of this study was to identify and develop aptamers against the hemagglutinin (HA) protein of influenza B virus.

An enriched DNA library with strong binding to the influenza B virus HA protein was obtained using magnetic bead systematic evolution of ligands by exponential enrichment technology after nine rounds of selection. Five candidate aptamers were identified by high-throughput sequencing. The aptamers were characterized using surface plasmon resonance and enzyme-linked immunosorbent assay techniques, and the aptamer exhibiting the highest affinity and specificity for the target protein was selected.

We screened and characterized five ssDNA aptamer sequences that bind to influenza B virus HA. Among these, aptamer sequence A573 exhibited the highest sensitivity and binding affinity for the target protein.

The novel aptamer sequences selected in this study have the potential to be used as biorecognition molecules for the development of aptamer sensors to detect influenza B virus.