Development of a Practical Method to Detect Noroviruses Contamination in Composite Meals |
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Authors: | Hiroyuki Saito Miho Toho Tomoyuki Tanaka Mamoru Noda |
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Institution: | 1.Akita Prefectural Research Center for Public Health and Environment,Akita,Japan;2.Fukui Prefectural Institute of Public Health and Environment Science,Fukui,Japan;3.Sakai City Institute of Public Health,Osaka,Japan;4.Division of Biomedical Food Research,National Institute of Health Sciences,Tokyo,Japan |
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Abstract: | Various methods to detect foodborne viruses including norovirus (NoV) in contaminated food have been developed. However, a practical method suitable for routine examination that can be applied for the detection of NoVs in oily, fatty, or emulsive food has not been established. In this study, we developed a new extraction and concentration method for detecting NoVs in contaminated composite meals. We spiked NoV-GI.4 or -GII.4 stool suspension into potato salad and stir-fried noodles. The food samples were suspended in homogenizing buffer and centrifuged to obtain a food emulsion. Then, anti-NoV-GI.4 or anti-NoV-GII.4 rabbit serum raised against recombinant virus-like particles or commercially available human gamma globulin and Staphylococcus aureus fixed with formalin as a source of protein A were added to the food emulsion. NoV-IgG-protein A-containing bacterial complexes were collected by centrifugation, and viral RNA was extracted. The detection limits of NoV RNA were 10–35 copies/g food for spiked NoVs in potato salad and stir-fried noodles. Human gamma globulin could also concentrate other NoV genotypes as well as other foodborne viruses, including sapovirus, hepatitis A virus, and adenovirus. This newly developed method can be used as to identify NoV contamination in composite foods and is also possibly applicable to other foodborne viruses. |
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