Norovirus, a member of the Caliciviridae family, is recognized as the principal cause of acute, non-bacterial gastroenteritis worldwide. It is estimated to cause 21 million cases of disease every year in the United States. Outbreaks are commonly identified in institutional as well as recreational settings, e.g. day care centers, hospitals, cruise ships and military settings. Norovirus has a very low infectious dose, has high resistance to traditional disinfection techniques, and multiple modes of transmission so early, point-of-care detection is crucial to control the spread of the disease.We have developed a new, rapid and sensitive lateral-flow assay technology using labeled phage particles that is potentially more rapid and economical than current diagnostics such RT-PCR and ELISA. Several affinity agents for norovirus detection including various antibody phage constructs as well as an anti-norovirus phage, expressing a peptide were systematically evaluated in this study. Capturing antibodies were adsorbed on lateral-flow membranes, before virus-like particles and detection phage were offered consecutively. The detection phage were modified with fluorescent dyes, e.g. Alexa-555 or reporter enzymes, such as horseradish peroxidase for a sensitive read-out utilizing a fluorescent reader, a scanner or by eye.

Norovirus detection using phage reporters in a lateral-flow assay

247th National Meeting (2014)

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