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Journal of Clinical Microbiology, July 2000, p. 2638-2642, Vol. 38, No. 7
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Detection of Herpes Simplex Virus DNA by Real-Time PCR

Harald H. Kessler,1,* Gerhard Mühlbauer,2 Beate Rinner,1 Evelyn Stelzl,1 Annemarie Berger,3 Hans-Wilhelm Dörr,3 Brigitte Santner,1 Egon Marth,1 and Holger Rabenau3

Institute of Hygiene, Karl-Franzens-University Graz, A-8010 Graz,1 and Roche Diagnostics GmbH, A-1211 Vienna,2 Austria, and Institute of Medical Virology, Johann-Wolfgang-Goethe-University Frankfurt, D-60596 Frankfurt am Main, Germany3

Received 27 January 2000/Returned for modification 3 April 2000/Accepted 8 May 2000

Molecular detection of herpes simplex virus (HSV) DNA is recognized as the reference standard assay method for the sensitive and specific diagnosis of central nervous system infections caused by HSV. In this study, a molecular assay based on real-time PCR on the LightCycler (LC) instrument was evaluated and compared with a home-brew molecular assay. The detection limit of the LC assay was determined with 10-fold dilutions of plasmid pS4 with the SalI restriction fragment of the DNA polymerase gene and with the First European Union Concerted Action HSV Proficiency Panel. A total of 59 cerebrospinal fluid (CSF) specimens were investigated for the comparative study. With plasmid pS4, the detection limit of the LC assay was found to be 104 copies per ml, i.e., 12.5 copies per run. When samples of the First European Union Concerted Action HSV Proficiency Panel were tested, 2 × 103 to 5 × 103 HSV type 1 genome equivalents (GE) per ml, i.e., 2.5 to 6.3 GE per run, could consistently be detected. There was a correlation between the LC assay and the home-brew assay in 55 of 59 specimens. In conclusion, the LC assay allows very rapid detection of HSV DNA in CSF. It was found to be laborsaving and showed sufficient sensitivity.

* Corresponding author. Mailing address: Molecular Diagnostics Laboratory, Institute of Hygiene, KF-University Graz, Universitaetsplatz 4, A-8010 Graz, Austria. Phone: 43(316)380-4363. Fax: 43(316)380-9649. E-mail: harald.kessler{at}kfunigraz.ac.at.

Journal of Clinical Microbiology, July 2000, p. 2638-2642, Vol. 38, No. 7
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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