Rapid method for SV40 titration.
SV40 titer is determined traditionally by the conventional plaque assay. Plaques appear after several rounds of infection and the assay takes around two weeks, which may delay research. A simpler assay was developed, based on detection of T-antigen in the infected cells by flow cytometry. Cells grown in 6-well plates are infected with serial dilutions of the viral stock, harvested 48h post-infection, stained and analyzed for T-antigen using a flow cytometer. The viral titer is calculated based on the percentage of T-antigen positive cells. The procedure is accomplished in 2 days. Unexpectedly we found that titers on different permissive African Green Monkey kidney cell lines were consistently different, suggesting variable susceptibility to SV40 infection. The method described, optimized for SV40 titration, may be adapted readily to other viruses.
Drayman
Nir
N
Department of Hematology, Hebrew University-Hadassah Medical School, Ein Kerem, Jerusalem, Israel.
Kler
Slava
S
Ben-nun-Shaul
Orly
O
Oppenheim
Ariella
A
D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000032:analysis; D002460:Cell Line; D002522:Chlorocebus aethiops; D005434:Flow Cytometry / Q000379:methods*; D013539:Simian virus 40 / Q000302:isolation & purification*; D013997:Time Factors; D019562:Viral Load / Q000379:methods*
D000952:Antigens, Polyomavirus Transforming