Generation and characterization of nucleic acid aptamers targeting the capsid P domain of a human norovirus GII.4 strain

•DNA aptamers were generated against the norovirus GII.4 P domain.•Aptamers showed broad reactivity with majority of a 14 strain capsid panel.•One aptamer showed at least low to moderate binding to all strains tested.•Aptamers bound dilutions of partially purified virus from clinical isolates.•Aptamers could concentrate norovirus from stool using magnetic capture. Human noroviruses (NoV) are the leading cause of acute viral gastroenteritis worldwide. Significant antigenic diversity of NoV strains has limited the availability of broadly reactive ligands for design of detection assays. The purpose of this work was to produce and characterize single stranded (ss)DNA aptamers with binding specificity to human NoV using an easily produced NoV target—the P domain protein. Aptamer selection was done using SELEX (Systematic Evolution of Ligands by EXponential enrichment) directed against an Escherichia coli-expressed and purified epidemic NoV GII.4 strain P domain. Two of six unique aptamers (designated M1 and M6-2) were chosen for characterization. Inclusivity testing using an enzyme-linked aptamer sorbent assay (ELASA) against a panel of 14 virus-like particles (VLPs) showed these aptamers had broad reactivity and exhibited strong binding to GI.7, GII.2, two GII.4 strains, and GII.7 VLPs. Aptamer M6-2 exhibited at least low to moderate binding to all VLPs tested. Aptamers significantly (p