DNA aptamers were developed against murine norovirus (MNV) using SELEX (Systematic Evolution of Ligands by EXponential enrichment). Nine rounds of SELEX led to the discovery of AG3, a promising aptamer with very high affinity for MNV as well as for lab-synthesized capsids of a common human norovirus (HuNoV) outbreak strain (GII.3). Using fluorescence anisotropy, AG3 was found to bind with MNV with affinity in the low picomolar range. The aptamer could cross-react with HuNoV though it was selected against MNV. As compared to a non-specific DNA control sequence, the norovirus-binding affinity of AG3 was about a million-fold higher. In further tests, the aptamer also showed nearly a million-fold higher affinity for the noroviruses than for the feline calicivirus (FCV), a virus similar in size and structure to noroviruses. AG3 was incorporated into a simple electrochemical sensor using a gold nanoparticle-modified screen-printed carbon electrode (GNPs-SPCE). The aptasensor could detect MNV with a limit of detection of approximately 180 virus particles, for possible on-site applications. The lead aptamer candidate and the aptasensor platform show promise for the rapid detection and identification of noroviruses in environmental and clinical samples.

Additional Metadata
Persistent URL dx.doi.org/10.1371/journal.pone.0079087
Journal PLoS ONE
Citation
Giamberardino, A. (Amanda), Labib, M. (Mahmoud), Hassan, E.M. (Eman M.), Tetro, J.A. (Jason A.), Springthorpe, S. (Susan), Sattar, S.A. (Syed A.), … DeRosa, M.C. (2013). Ultrasensitive norovirus detection using DNA aptasensor technology. PLoS ONE, 8(11). doi:10.1371/journal.pone.0079087