Culturing, storage, and quantification of rotaviruses.
Group A rotaviruses (RVs) infect the young of numerous animal species and cause acute gastroenteritis. Cultivation of animal and human RVs in cells requires proteolytic activation of the viral attachment protein using trypsin. Continuous cell lines, such as rhesus monkey kidney cells, as well as primary monkey kidney cells, are routinely used for the growth and characterization of RVs. Isolation and cultivation of human RVs from clinical fecal specimens is difficult and adaptation to growth in vitro requires multiple rounds of passage in primary cells. Following growth, RV stocks can be purified by centrifugation, if required, and quantified using plaque assay or fluorescence focus assay. This unit describes easily applicable procedures for the culturing, storage, and quantification of RVs.
Arnold
Michelle
M
Laboratory of Infectious Diseases, NIAID/NIH, Bethesda, Maryland, USA.
Patton
John T
JT
McDonald
Sarah M
SM
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D002460:Cell Line; D005612:Freeze Drying / Q000379:methods; D006801:Humans; D012401:Rotavirus / Q000737:chemistry / Q000254:growth & development*; D012400:Rotavirus Infections / Q000821:virology; D014776:Virus Cultivation / Q000379:methods*