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Riley, Jean M. (U.S. Army Biological Laboratories, Fort Detrick, Frederick, Md.), William C. Patrick, III, and William E. Campbell, Jr. Estimation of titer of Venezuelan equine encephalomyelitis virus preparations from a single-dilution assay. J. Bacteriol. 85:1256-1260. 1963.-When suspensions of Venezuelan equine encephalomyelitis (VEE) virus were injected intracerebrally into groups of mice, a nearly linear relationship was observed between the concentration of the virus injected and the mean reciprocal time-to-death of the mice. A total of 91 VEE preparations were assayed in duplicate, and, by plotting the relationship between the reciprocal time-to-death for mice given the 10(-6) dilution of virus and the micld(50) (mouse intracerebral challenge, ld(50) response) values for the virus preparations, a reference curve was established. Using this reference curve, it was possible to estimate directly the ld(50) values of virus suspensions of unknown concentration from the mean reciprocal time-to-death of a group of mice injected with a single dilution. In this work, the number of mice used was reduced by 62.5%, the titrations were complete in 3 to 5 days compared with the usual 10 to 14 days, three to four times as many assays could be done in a day, and no assays had to be repeated since end points were not missed. The precision of the single-dilution assay compared favorably with that of the ld(50) titration.

ESTIMATION OF TITER OF VENEZUELAN EQUINE ENCEPHALOMYELITIS VIRUS PREPARATIONS FROM A SINGLE-DILUTION ASSAY