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Journal of Clinical Microbiology, February 2001, p. 460-463, Vol. 39, No. 2
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.2.460-463.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Sensitive Detection of Ehrlichia chaffeensis in Cell Culture, Blood, and Tick Specimens by Reverse Transcription-PCR

Suleyman Felek,1,dagger Ahmet Unver,1 Roger W. Stich,2 and Yasuko Rikihisa1,*

Department of Veterinary Biosciences1 and Department of Veterinary Preventive Medicine,2 College of Veterinary Medicine, The Ohio State University, Columbus, Ohio 43210-1093

Received 25 July 2000/Returned for modification 26 October 2000/Accepted 8 November 2000

Ehrlichia chaffeensis is an obligatory intracellular bacterium of monocytes and macrophages and the etiologic agent of human monocytic ehrlichiosis, an emerging zoonosis. The Lone Star tick (Amblyomma americanum) has been implicated as the primary vector of E. chaffeensis. The present study examined the sensitivity of the nested reverse transcription (RT)-PCR based on the 16S rRNA gene relative to that of the nested PCR for detection of E. chaffeensis in infected DH82 cells, experimentally infected dog peripheral blood mononuclear cells, or experimentally infected A. americanum tick samples. The RT-PCR was found to be approximately 100 times more sensitive than the PCR for detection of E. chaffeensis regardless of the nature of the specimens. Thus, this RT-PCR is useful for detection of E. chaffeensis when a high sensitivity is required. Positive results by RT-PCR also imply the presence of viable pathogens. This is the first demonstration of RNA of E. chaffeensis in infected blood and acquisition-fed male, nymphal, and larval A. americanum ticks.

* Corresponding author. Mailing address: Department of Veterinary Biosciences, College of Veterinary Medicine, The Ohio State University, 1925 Coffey Rd., Columbus, OH 43210-1093. Phone: (614) 292-5661. Fax: (614) 292-6473. E-mail: rikihisa.1{at}osu.edu.

dagger Present address: Department of Clinical Microbiology and Infectious Diseases, School of Medicine, Firat University, Elazig, Turkey 23119.

Journal of Clinical Microbiology, February 2001, p. 460-463, Vol. 39, No. 2
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.2.460-463.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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