Key result
Single stranded DNA aptamers targeting the human norovirus P domain exhibited broad reactivity to diverse virus-like particles and significantly bound virus in clinical stool specimens (p<0.05).
This study demonstrates that the human norovirus P domain protein is a functional target for selecting nucleic acid aptamers that specifically bind and broadly recognize diverse human norovirus strains.
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May enable aptamer-based NoV detection; leaves open clinical validation across strains and settings.
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<0.05) bound virus in partially purified GII.4 New Orleans outbreak stool specimens as demonstrated by ELASA and aptamer magnetic capture (AMC) followed by RT-qPCR. This is the first demonstration of human NoV P domain protein as a functional target for the selection of nucleic acid aptamers that specifically bind and broadly recognize diverse human NoV strains.
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Moore et al. (2015) studied Human norovirus. Single stranded (ss)DNA aptamers targeting NoV P domain was evaluated on Binding specificity and reactivity to human NoV VLPs and virus in stool specimens (p=<0.05). Single stranded DNA aptamers targeting the human norovirus P domain exhibited broad reactivity to diverse virus-like particles and significantly bound virus in clinical stool specimens (p<0.05).
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