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Antimicrobial Agents and Chemotherapy, September 2002, p. 2943-2947, Vol. 46, No. 9
0066-4804/02/$04.00+0     DOI: 10.1128/AAC.46.9.2943-2947.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Application of Real-Time PCR for Determination of Antiviral Drug Susceptibility of Herpes Simplex Virus

Ruzena Stránská,* Anton M. van Loon, Merjo Polman, and Rob Schuurman

Department of Virology, Eijkman-Winkler Center, University Medical Center Utrecht, The Netherlands

Received 13 December 2001/ Returned for modification 12 March 2002/ Accepted 30 May 2002

A quantitative real-time PCR (TaqMan) assay was developed for determination of antiviral drug susceptibility of herpes simplex virus (HSV). After short-time culture of the virus, the antiviral drug susceptibility of HSV isolates for acyclovir (ACV) was determined by measuring the reduction of the HSV type 1 (HSV-1) DNA levels in culture supernatants using real-time PCR. The 50% inhibitory concentration was reported as the concentration of antiviral drug that reduced the number of HSV-1 DNA copies by 50%. A total of 15 well-characterized ACV-sensitive or -resistant strains and clinical isolates were used for assay evaluation. The new assay with real-time PCR readout permitted rapid (3 days), objective, and reproducible determination of HSV-1 drug susceptibilities with no need for stringent control of initial multiplicity of infection. Furthermore, the real-time PCR assay results showed good correlation (r = 0.86) with those for the plaque reduction assay. In conclusion, the real-time PCR assay described here is a suitable quantitative method for determination of antiviral susceptibility of HSV-1, amenable for use in the routine diagnostic virology laboratory.

* Corresponding author. Mailing address: Department of Virology, University Medical Center Utrecht, Heidelberglaan 100, 3584 CX, Utrecht, The Netherlands. Phone: 31 30 250 6526. Fax: 31 30 250 5426. E-mail: r.stranska{at}lab.azu.nl.

Antimicrobial Agents and Chemotherapy, September 2002, p. 2943-2947, Vol. 46, No. 9
0066-4804/02/$04.00+0     DOI: 10.1128/AAC.46.9.2943-2947.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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  • Mace, M., Manichanh, C., Bonnafous, P., Precigout, S., Boutolleau, D., Gautheret-Dejean, A., Agut, H. (2003). Real-Time PCR as a Versatile Tool for Investigating the Susceptibility of Human Herpesvirus 6 to Antiviral Agents. Antimicrob. Agents Chemother. 47: 3021-3024 [Abstract] [Full Text]  
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