Culturing, storage, and quantification of rotaviruses

Area of Science:

• Virology
• Microbiology
• Cell Biology

Background:

• Group A rotaviruses (RVs) are significant pathogens causing acute gastroenteritis in young animals.
• Cultivation of RVs in cell cultures necessitates proteolytic activation of the viral attachment protein with trypsin.
• Standard methods involve continuous or primary monkey kidney cells for RV growth and characterization.

Purpose of the Study:

• To describe easily applicable procedures for the culturing, storage, and quantification of Group A rotaviruses (RVs).
• To provide a standardized protocol for researchers working with animal and human rotaviruses.
• To facilitate the growth and characterization of RVs in vitro.

Main Methods:

• Culturing of RVs in continuous cell lines (e.g., rhesus monkey kidney cells) or primary monkey kidney cells.
• Proteolytic activation of the viral attachment protein using trypsin.
• Purification of RV stocks via centrifugation and quantification using plaque assay or fluorescence focus assay.

Main Results:

• Established protocols for the successful in vitro cultivation of Group A rotaviruses.
• Demonstrated methods for the storage and preservation of infectious RV stocks.
• Validated techniques for accurate RV quantification essential for experimental design.

Conclusions:

• The described procedures offer a straightforward approach for culturing, storing, and quantifying Group A rotaviruses.
• These methods are crucial for advancing research into RV pathogenesis and developing effective countermeasures.
• Standardized protocols enhance reproducibility and facilitate the study of diverse RV strains.