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Highly sensitive ELISA for the serological detection of murine rotavirus EDIM based on its major immunogen VP6

 
: Fingas, Felix; Rückner, Antje; Heenemann, Kristin; Volke, Daniela; Sieg, Michael; Bielefeld, Petra; Grunwald, Thomas; Vahlenkamp, Thomas W.; Hassert, Rayk; Hoffmann, Ralf

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Journal of virological methods 262 (2018), pp.72-78
ISSN: 0166-0934
ISSN: 1879-0984
English
Journal Article
Fraunhofer IZI ()
rotavirus EDIM; ELISA; VP6; recombinant protein; FELASA

Abstract
Precise health monitoring of laboratory animals is a critical factor for surveillance and accuracy of animal experiments. Rotavirus epizootic diarrhea of infant mice (EDIM) leads to infections in mice that can influence animal studies, e.g., by altering the intestinal physiology. Thus, the aim of this study was establishing a highly sensitive and specific ELISA for the serological detection of EDIM infections in rodents. First, virus proteins were separated by SDS-PAGE and immunogenic proteins were visualized by immunoblotting and identified after ingeldigestion by tandem mass spectrometry. Subsequently, the major immunogen VP6 (virus protein 6) was expressed in Escherichia coli in high yields, purified by affinity chromatography, and used to establish an indirect ELISA. The diagnostic sensitivity and specificity were both above 99 % and the selectivity better than 98.7 % for animals infected by other pathogens listed by the Federation of Laboratory Animal Science Associations. Importantly, the Strep-rVP6-His-ELISA was more sensitive than a commercial virus-based ELISA and is a time and cost-efficient complement to EDIM-specific immune-fluorescence assays. In conclusion, the assay can improve health monitoring by reducing the risk of missed EDIM infections in animal housing facilities, thereby improving animal welfare, reliability of animal studies, and protection of precious mice breeds.

: http://publica.fraunhofer.de/documents/N-515470.html