title: Surface plasmon resonance biosensor for detection of feline calicivirus, a surrogate for norovirus.
abstract: The human noroviruses are the most common non-bacterial cause of gastroenteritis and are responsible for as much as 50% of all gastroenteritis outbreaks worldwide. Norovirus (NoV), a single stranded RNA virus, is highly contagious with an infectious dose of less than 100 viral particles. While techniques exist for the identification of NoV, the lack of a reliable cell culture system, NoV genetic variability, and time-consuming sample preparation steps required to isolate the virus (or its genome) prior to molecular based methods has hindered rapid virus detection. To better protect the public from virus-contaminated food and enable better detection in clinical and environmental samples, sensitive and selective methods with simple sample preparation are needed. Surface plasmon resonance (SPR) biosensors represent an emerging detection platform, and this approach has been applied to the rapid detection of foodborne small molecule toxins, protein toxins, and bacteria. This analytical technique, however, has yet to be fully investigated for rapid virus detection, especially for intact viral particles extracted from food matrices. For this study, the culturable, non-human pathogen feline calicivirus (FCV), which has similar morphology and is genetically related to NoV, was chosen as a surrogate virus for designing and evaluating an SPR assay. An antibody-based assay was performed by first immobilizing anti-FCV to an SPR chip surface and then directly measuring virus binding and subsequent secondary antibody binding. The resulting biosensor directly detected intact FCV particles with limits of detection of approximately 10(4)TCID50FCV/mL from purified cell culture lysates. In addition, intact virus detection in FCV-spiked oyster matrix was possible when using a simple extraction procedure and employing a secondary antibody to FCV for quantitation. The results from these preliminary studies show promise for the development of a rapid assay for detecting intact viruses, such as NoV, using an SPR biosensor. While the current level of sensitivity achieved with this SPR biosensor may be more applicable to virus detection in clinical specimens, broader application and increased sensitivity of this method for foodborne viruses may be achieved when performed in conjunction with efficient virus extraction and concentration methods.
authors: U.S. Food and Drug Administration, Center for Food Safety and Applied Nutrition, Office of Regulatory Science, College Park, MD 20740, USA. betsy.yakes@fda.hhs.gov
authors: Betsy Jean
authors: BJ
authors: Yakes
authors: Efstathia
authors: E
authors: Papafragkou
authors: Stephen M
authors: SM
authors: Conrad
authors: John D
authors: JD
authors: Neill
authors: Julia F
authors: JF
authors: Ridpath
authors: William
authors: W
authors: Burkhardt
authors: Michael
authors: M
authors: Kulka
authors: Stacey L
authors: SL
authors: Degrasse
mesh_terms: D015374:Biosensing Techniques; D017927:Calicivirus, Feline / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D059022:Food Safety / Q000379:methods*; D005759:Gastroenteritis / Q000517:prevention & control / Q000821:virology; D057230:Limit of Detection; D029322:Norovirus / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D020349:Surface Plasmon Resonance