Neurovirulence of individual plaque stocks of Herpes simplex virus type 2 strain HG 52.

The virulence of ten independent plaque stocks of HSV-2 strain HG 52 was determined by intracranial inoculation of 3 week old BALB/c mice. The ten stocks exhibited a range of LD50 which could be grouped into three classes; high (less than 10(3) pfu/mouse), intermediate (10(3)-10(4) pfu/mouse) and low (greater than 10(5) pfu/mouse). Analysis of the ten stocks using six different restriction endonucleases showed no significant variation in the size and distribution of fragments. Isolates from the brains of dead mice had restriction enzyme profiles indistinguishable from the initial infecting virus. The pfu:particle ratios of the virus stocks were comparable. There were no significant differences between the single growth cycle characteristics in vitro of the parental HG 52 and the various plaque purified stocks. In vivo after intracranial inoculation the low virulence stocks grew poorly while those of high virulence grew well. Plaques were picked from the high and low virulence stocks after passage five times in vitro and re-assayed for virulence. In some cases there were differences in the LD50s of passaged plaque isolates from that of parental stock. The restriction enzyme profiles of the passaged plaque isolates remained unchanged. The analysis has clearly demonstrated virulence heterogeneity within a single HSV-2 virus stock.