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Journal of Clinical Microbiology, July 2002, p. 2445-2451, Vol. 40, No. 7
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.7.2445-2451.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Real-Time PCR for Quantification of Human Herpesvirus 6 DNA from Lymph Nodes and Saliva

Departments of Virology,¹ Pathology,² Clinical Haematology, Limoges University Teaching Hospital, Limoges, France³

Received 13 November 2001/ Returned for modification 20 February 2002/ Accepted 5 April 2002

A real-time quantitative PCR assay has been developed to measure human herpesvirus 6 (HHV-6) DNA in biological specimens. The assay sensitivity was 10 copies of DNA per well, with a linear dynamic range of 10 to 10⁷ copies of HHV-6 DNA. Intra- and interassay variations were, respectively, 0.88 and 0.8% for samples containing 10² DNA copies, 0.99 and 0.96% for samples containing 10⁴ copies, and 0.76 and 0.9% for samples containing 10⁶ copies. Among 34 saliva samples from healthy subjects, 26 were found to contain HHV-6 DNA (76.5%; median, 23,870 copies/ml), and following a single freeze-thaw cycle, 25 of the same samples were found to be positive for HHV-6 DNA, although at a statistically significantly lower concentration (median, 3,497 copies/ml). The assay enabled detection of HHV-6 DNA in lymph node biopsies from patients with Hodgkin's disease (HD) (13 of 37 patients [35.1%]), B-cell neoplasms (8 of 36 patients [22.2%]), and T- or NK-cell neoplasms (3 of 13 patients [23.1%]), with concentrations ranging from 100 to 864,640 HHV-6 copies per µg of DNA (HHV-6B being found in every case except two). All HD patients infected with HHV-6 presented clinically with the nodular sclerosis subtype of HD. The real-time quantitative PCR assay developed here was simple to perform and was sensitive over a wide range of HHV-6 concentrations. It therefore appears to be of potential value in clinical investigation or diagnosis of HHV-6 infection.

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