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Journal of Clinical Microbiology, May 1999, p. 1329-1331, Vol. 37, No. 5
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Quantitative Real-Time PCR for Detection of Members of the Ehrlichia phagocytophila Genogroup in Host Animals and Ixodes ricinus Ticks

Nicola Pusterla,1,* Jon B. Huder,2 Christian M. Leutenegger,1 Ueli Braun,1 John E. Madigan,3 and Hans Lutz1

Department of Veterinary Internal Medicine, University of Zurich, CH-8057 Zurich,1 and Swiss National Center for Retroviruses, University of Zurich, CH-8044 Zurich,2 Switzerland, and Department of Medicine and Epidemiology, School of Veterinary Medicine, University of California, Davis, California 956163

Received 27 October 1998/Returned for modification 7 December 1998/Accepted 26 January 1999

A TaqMan PCR was established for identification and quantitation of members of the Ehrlichia phagocytophila group in experimentally infected cows and in Ixodes ricinus ticks. The TaqMan PCR identified a 106-bp section of the 16S rRNA gene by use of a specific fluorogenic probe and two primers. This technique was specific for members of the E. phagocytophila group, which include E. phagocytophila, Ehrlichia equi, and the agent of human granulocytic ehrlichiosis. The TaqMan system identified 10 copies of a cloned section of the 16S rRNA gene of E. phagocytophila. The sensitivity and specificity of the TaqMan PCR were similar to those of conventional nested PCR. The numbers of ehrlichiae in leukocytes of the two cows experimentally infected with E. phagocytophila were measured daily by TaqMan PCR and had a course similar to that of the percentages of infected leukocytes determined daily by light microscopy. The prevalence of infected free-living ticks, which were collected from areas where bovine ehrlichiosis is endemic and from regions with sporadic occurrences of granulocytic ehrlichiosis in dogs and horses, was identical as determined by nested PCR and TaqMan PCR.

* Corresponding author. Present address: School of Veterinary Medicine, University of California, Davis, CA 95616. Phone: (530) 752-2371. Fax: (530) 752-0414. E-mail: npusterla{at}ucdavis.edu.

Journal of Clinical Microbiology, May 1999, p. 1329-1331, Vol. 37, No. 5
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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