A real-time quantitative polymerase chain reaction was utilized to study the in vivo replication of Marek's disease vaccine viruses and of virulent oncogenic strains. In the first of four experiments, the growth of the herpes virus of turkeys (HVT) vaccine was detectable in various organs of infected chicken embryos, with the highest viral loads being present in the spleen. No evidence was obtained for replication of serotype-1 Marek's disease viruses in embryos. In the second experiment, viral loads were measured in several organs of chickens after administration of the Rispens and HVT vaccines immediately after hatching. Lowest levels were noted for the Rispens strain after 1 to 8 weeks. By contrast, HVT vaccine grew well in all tested organs, with the highest loads being present in the spleen. Highest loads were observed in unvaccinated birds after challenge with the highly virulent strain MPF57 at day 8, especially in lymphoid organs. A positive relationship was observed between viral load and clinical signs, including tumour formation. In a third study, viral loads were measured in the organs of chickens administered the Rispens vaccine on the day of hatch and challenged at day 8 with MPF57. High levels of clinical protection were afforded against MPF57 by the Rispens vaccine but, in confirmation of earlier findings, sterilizing immunity was not induced. In a fourth study, two experiments were conducted--in which viral loads were measured after challenge of chickens vaccinated with HVT in ovo or at day 1 after hatching. Similar protection was achieved in birds vaccinated in ovo on embryonic days 11 and 17, although protection was slightly, but not significantly, lower than for birds vaccinated at day 1.