The use of real-time PCR has been described previously for analysing both the replication kinetics and antiviral susceptibility of human herpesvirus 6 in MT4 cells. It is now reported that viral DNA persists in infected cell culture long after the end of lytic virus replication. Consequently, high levels of DNA may correspond to an absence of infectivity and late readout occurring after the exponential phase of virus growth may lead to misinterpretation of the results of susceptibility assays. These limitations must be borne in mind when using real-time PCR.