Quantitation of Friend spleen focus-forming virus by a nine-day 59Fe assay

A previously described 3-day 59Fe assay for quantitation of Friend spleen focus-forming virus has been modified to produce a 200-fold more sensitive 9-day 59Fe assay. A characterization of this assay is reported here. Male BALB/c mice received intravenous injections of appropriately diluted Friend polycythemia virus (FVP); control mice received virus diluent. All mice were allowed food and water ad libitum for 6 days, and on day 6 after virus injection were fasted by removal of food but not water. On day 3 of the fast (the 9th day after virus injection) each mouse received an intraperitoneal injection of 1 muCi of 59Fe. Six hours later the mice were sacrificed and the splenic radioactivity was determined. The percent splenic incorporation of 59Fe was directly related to the logarithm of spleen focus-forming units (SFFU) of FVP injected in a range of approximately 25 to 1,000 SFFU. Using a standard FVP preparation in a dose range of 25 to 1,000 SFFU, it was possible to determine the SFFU titers of unknown samples by extrapolation of the percent splenic 59Fe incorporation to the logarithm of SFFU. SFFU titers obtained by the 9-day 59Fe assay were similar to those obtained by the enumerative-response assay. Advantages of the 9-day 59Fe assay over the enumerative-response assay include a 50-fold greater virus dose range, an easier and a more objective counting procedure, and a reduced coefficient of variation.