Ultrasensitive Norovirus Detection Using DNA Aptasensor Technology |
| |
| Authors: | Amanda Giamberardino Mahmoud Labib Eman M Hassan Jason A Tetro Susan Springthorpe Syed A Sattar Maxim V Berezovski Maria C DeRosa |
| |
| Institution: | 1. Department of Chemistry, Carleton University, Steacie Building, Ottawa, Ontario, Canada.; 2. Department of Chemistry, University of Ottawa, Ottawa, Ontario, Canada.; 3. Centre for Research on Environmental Microbiology, University of Ottawa, Ottawa, Ontario, Canada.; National Institute of Allergy and Infectious Diseases, United States of America, |
| |
| Abstract: | 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. |
| |
| Keywords: | |
|
|
