Preparation of antiserum against rotavirus for use in diagnostic kits based on ELISA and immunofluorescence

Introduction and Objectives: Rotavirus is a major cause of severe watery diarrhea, vomiting, fever, and abdominal pain. This virus can spread easily among infants and young children. Children who get rotavirus disease can become dehydrated and may need to be hospitalized. It is not easy to distinguish gastroenteritis caused by rotavirus from other enteric pathogens. Therefore, establishing an adequate diagnosis requires testing of fecal specimens with commercially available assays like electron microscopy, immunofluorescence and Elisa. The purpose of this study was to provide antiserum against rotavirus antigens for use in ELISA and immunofluorescence kits.Materials and Methods: Inactivated virus was injected intravenously into New Zealand White Rabbit on a weekly basis. Blood collection was performed 7 days following last injection. Antibody was purified with different methods like Ammonium sulfate precipitation, Tangential flow filtration and Ion Exchange chromatography.Results: Results showed adequate quality and quantity of produced antibody. The purity was confirmed by SDS-Page electrophoresis and protein concentration also checked with Bradford method. Purified antibody was used in Elisa after biotinylation. Clinical specimens were checked by designed Elisa kit. Conclusion: Using antibody and biotinylated antibody in Elisa kit showed acceptable sensitivity and specificity and could provide a convenient, rapid and reliable diagnostic method for rotavirus infections.

کلیدواژه ها:

Rotavirus ، Antiserum ، Elisa ، Immunofluorescence.