alexa Comparison of ELISA attachment and infectivity assays for murine norovirus.
Chemical Engineering

Chemical Engineering

Journal of Analytical & Bioanalytical Techniques

Author(s): Hirneisen KA, Kniel KE

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Abstract Human noroviruses cause approximately 58\% of foodborne illnesses in the USA. Recent studies have shown norovirus attachment to the carbohydrates moieties of host cellular receptors. Using murine norovirus (MNV) as a surrogate, an ELISA method was utilized to assess attachment through binding to host cell receptors; MNV attachment was correlated to infectivity determined by plaque assay. ELISA plates were coated with porcine gastric mucin and untreated, heat-, high pressure-, ozone- and UV-treated MNV was added followed by monoclonal anti-MNV IgG antibody. The average OD(405) of MNV-containing wells were divided by negative control wells and expressed as the 'P/N ratio'; values ≥2 were considered positive. Infectivity of MNV following heat and HPP treatments was determined using the plaque assay. Heat-treated MNV attachment decreased significantly with decreasing viral infectivity whereby the P/N ratio was <2 after treatment at 80 and 100°C for 5 min which correlated with a non-intact capsid as shown by RNase treatment. No significant difference in attachment was observed for pressure-, ozone- and UV-treated MNV. These findings suggest potentially different effects on the viral capsid due to different food processing methods. Copyright © 2012 Elsevier B.V. All rights reserved. This article was published in J Virol Methods and referenced in Journal of Analytical & Bioanalytical Techniques

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