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Plaque quantitation and virus neutralization assays for human astroviruses

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Summary

Astroviruses, 28 nm-diameter, RNA-containing viruses which have been implicated in gastroenteritis can be cultivated in cell cultures containing trypsin, but do not show distinguishable cytopathic effects. However, with the 5 known astrovirus serotypes which we have been able to cultivate, 3 (types 1, 2, and 5) formed well-defined plaques in LLCMK2 cell cultures under an agar overlay containing trypsin. A virus neutralization assay based on plaque reduction was applied to these 3 serotypes. It was found that rabbit antisera prepared against individual serotypes neutralized virus type-specifically, and no cross-neutralization titers were obtained with any of the antisera to the 5 astrovirus serotypes. The type-specific neutralization observed agreed with the specificities seen by immunofluorescence (IF), whereas ELISA tests with the same antisera show cross-reactivity among all 5 serotypes. There was no virus neutralization detected with astrovirus monoclonal antibodies which were reactive with 5 serotypes by ELISA and IF. The results we have obtained permit quantitative techniques to be applied to epidemiological and biological studies of the human astroviruses.

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Hudson, R.W., Herrmann, J.E. & Blacklow, N.R. Plaque quantitation and virus neutralization assays for human astroviruses. Archives of Virology 108, 33–38 (1989). https://doi.org/10.1007/BF01313740

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  • DOI: https://doi.org/10.1007/BF01313740

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