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Journal of Clinical Microbiology, November 2001, p. 3982-3986, Vol. 39, No. 11
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.11.3982-3986.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Western Blot Analysis of Sera Reactive to Human Monocytic Ehrlichiosis and Human Granulocytic Ehrlichiosis Agents

Ahmet Unver,1,dagger Suleyman Felek,1 Christopher D. Paddock,2 Ning Zhi,1 Harold W. Horowitz,3 Gary P. Wormser,3 Louis C. Cullman,4,Dagger and Yasuko Rikihisa1,*

Department of Veterinary Biosciences, College of Veterinary Medicine, The Ohio State University, Columbus, Columbus, Ohio 43210-10931; Viral and Rickettsial Zoonoses Branch, Centers for Disease Control and Prevention, Atlanta, Georgia 303332; Division of Infectious Diseases, Department of Medicine, New York Medical College, Valhalla, New York 105953; and MRL Reference Laboratory, Cypress, California 906304

Received 21 May 2001/Returned for modification 30 July 2001/Accepted 3 September 2001

Laboratory diagnosis of human ehrlichioses is routinely made by an indirect immunofluorescence assay (IFA) using cultured ehrlichia-infected whole cells as antigen. Concern has been raised that incorrect diagnoses of human monocytic ehrlichiosis (HME) or human granulocytic ehrlichiosis (HGE) may be made on the basis of serologic cross-reactivity between Ehrlichia chaffeensis and the agent of HGE. The present study examined whether two recombinant major outer membrane proteins, rP30 and rP44, that were previously shown to be sensitive and specific serodiagnostic antigens for HME and HGE, respectively, could be used to discriminate IFA dually reacting sera. Thirteen dually IFA-reactive sera, three sera that were IFA positive only with E. chaffeensis, and three sera that were IFA positive only with the HGE agent were examined by Western immunoblot analysis using purified whole organisms and recombinant proteins as antigens. All 16 E. chaffeensis IFA-positive sera reacted with rP30. However, none of these sera reacted with rP44, regardless of IFA reactivity with the HGE agent. The three HGE-agent-only IFA-positive sera reacted only with rP44, not with rP30. Western immunoblotting using purified E. chaffeensis and the HGE agent as antigens suggested that heat shock and other proteins, but not major outer membrane proteins, cross-react between the two organisms. Therefore, Western immunoblot analysis using rP44 and rP30 may be useful in discriminating dually HME and HGE IFA-reactive sera.

* 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 Microbiology, Faculty of Veterinary Medicine, Kafkas University, Kars, Turkey.

Dagger Present address: Oppenheimer Wolff and Donnelly, Newport Beach, CA 92660.

Journal of Clinical Microbiology, November 2001, p. 3982-3986, Vol. 39, No. 11
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.11.3982-3986.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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Copyright © 2001 by the American Society for Microbiology. All rights reserved.