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Journal of Clinical Microbiology, February 2002, p. 519-523, Vol. 40, No. 2
0095-1137/01/$04.00+0     DOI: 10.1128/JCM.40.2.519-523.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Quantification of Feline Herpesvirus 1 DNA in Ocular Fluid Samples of Clinically Diseased Cats by Real-Time TaqMan PCR

A. Vögtlin,1 C. Fraefel,1 S. Albini,1 C. M. Leutenegger,2 E. Schraner,3 B. Spiess,4 H. Lutz,2 and M. Ackermann1*

Institutes of Virology,1 AnatomyDepartments of,3 Internal Veterinary Medicine,2 Veterinary Surgery, Veterinary Medical Faculty, University of Zurich, Zurich, Switzerland4

Received 22 June 2001/ Returned for modification 9 October 2001/ Accepted 4 November 2001

A fluorogenic PCR was established for the quantification of feline herpesvirus 1 (FeHV-1) DNA in ocular fluid samples of clinically diseased cats. The new assay was specific for FeHV-1 and sensitive. The 100% detection rate ranged from 0.6 to 6 50% tissue culture infective doses per sample. When spiked samples with known quantities of virus were used, infectious virus titers and quantification of viral DNA by PCR correlated to each other in a linear fashion (R2 = 0.9858) over a range of 4 orders of magnitude. Within this range, it was possible to calculate the FeHV-1 DNA content from a given infectious dose, and vice versa. The new diagnostic procedure was applied to ocular fluid samples from cats experimentally infected with FeHV-1 and specific FeHV-1-free cats. A good correlation between virus titer and quantitative PCR was observed, although only early in infection. In a second stage, the titer of infectious virus collapsed, while the PCR signal remained high. A constantly decreasing PCR signal accompanied by negative virus isolation was characteristic for a final stage of the infection. Finally, clinical samples from 20 cats that were suspected to suffer from FeHV-1 infection were analyzed. By comparing virus titers and quantitative PCR signals, it was possible to determine the current stage of the ongoing infection. Based on these findings, comparison of the results of consecutive samples allows the tracking of the course of the infection. Therefore, the new method combines the advantages of the two previously established conventional methods, qualitative PCR and virus isolation and titration.

* Corresponding author. Mailing address: Institute of Virology, Veterinary Medical Faculty, University of Zurich, Winterthurerstrasse 266a, CH-8057 Zurich, Switzerland. Phone: 41 1 635 87 01. Fax: 41 1 635 89 11. E-mail: ma{at}vetvir.unizh.ch.

Journal of Clinical Microbiology, February 2002, p. 519-523, Vol. 40, No. 2
0095-1137/01/$04.00+0     DOI: 10.1128/JCM.40.2.519-523.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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