Detection of herpes viruses in clinical samples using real-time PCR

Herpes viruses including cytomegalovirus, varicella tester and herpes simpl ex are an important cause of morbidity and mortality, especially in immunoc ompromised patients. Real-time PCR assays were developed with the aim of in troducing a rapid and sensitive test to replace culture, and as a surveilla nce system for high-risk patients. The assays were optimised using cell cul ture derived material, and the sensitivity ascertained using cloned product before applying to extracted and non-extracted clinical samples. The sensi tivity was between 1-100 virus copies with increased sensitivity to detect less than 10 copies possible when an initial round of amplification was car ried out using external primers. Results were available within four hours o f receipt compared with a median of 4.4 days for culture and immunofluoresc ence. Real-time PCR was found to be a sensitive and rapid method of detecti ng these viruses and will be a valuable tool for the surveillance of immuno suppressed patients. (C) 2001 Elsevier Science B.V. All rights reserved.