title: Comparison of ELISA attachment and infectivity assays for murine norovirus.
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.
authors: Department of Animal and Food Sciences, University of Delaware, Newark, DE 19716, USA.
authors: Kirsten A
authors: KA
authors: Hirneisen
authors: Kalmia E
authors: KE
authors: Kniel
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D000914:Antibodies, Viral / Q000378:metabolism; D004202:Disinfectants / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006358:Hot Temperature; D006874:Hydrostatic Pressure; D007074:Immunoglobulin G / Q000378:metabolism; D051379:Mice; D050296:Microbial Viability / Q000187:drug effects / Q000528:radiation effects; D009077:Mucins / Q000378:metabolism; D029322:Norovirus / Q000187:drug effects / Q000502:physiology* / Q000528:radiation effects; D011485:Protein Binding; D011991:Receptors, Virus / Q000378:metabolism; D013552:Swine; D010948:Viral Plaque Assay / Q000379:methods; D014773:Virology / Q000379:methods*; D053585:Virus Attachment
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D004202:Disinfectants; D007074:Immunoglobulin G; D009077:Mucins; D011991:Receptors, Virus