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Clinical and Diagnostic Laboratory Immunology, July 2003, p. 520-524, Vol. 10, No. 4
1071-412X/03/$08.00+0     DOI: 10.1128/CDLI.10.4.520-524.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Characterization of the Major Antigenic Protein 2 of Ehrlichia canis and Ehrlichia chaffeensis and Its Application for Serodiagnosis of Ehrlichiosis

Tamece T. Knowles,¹ A. Rick Alleman,^2* Heather L. Sorenson,² David C. Marciano,¹ Edward B. Breitschwerdt,³ Shimon Harrus,⁴ Anthony F. Barbet,¹ and Myriam Bélanger^1,

Departments of Pathobiology,¹ Physiological Sciences, College of Veterinary Medicine, University of Florida, Gainesville, Florida 32610,² Department of Companion Animal and Special Species Medicine, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina 27606,³ School of Veterinary Medicine, The Hebrew University of Jerusalem, Rehovot, Israel 76100⁴

Received 11 September 2002/ Returned for modification 27 November 2002/ Accepted 11 March 2003

Canine monocytic ehrlichiosis, caused by Ehrlichia canis or Ehrlichia chaffeensis, can result in clinical disease in naturally infected animals. Coinfections with these agents may be common in certain areas of endemicity. Currently, a species-specific method for serological diagnosis of monocytic ehrlichiosis is not available. Previously, we developed two indirect enzyme-linked immunosorbent assays (ELISAs) using the major antigenic protein 2 (MAP2) of E. chaffeensis and E. canis. In this study, we further characterized the conservation of MAP2 among various geographic isolates of each organism and determined if the recombinant MAP2 (rMAP2) of E. chaffeensis would cross-react with E. canis-infected dog sera. Genomic Southern blot analysis using digoxigenin-labeled species-specific probes suggested that map2 is a single-copy gene in both Ehrlichia species. Sequences of the single map2 genes of seven geographically different isolates of E. chaffeensis and five isolates of E. canis are highly conserved among the various isolates of each respective ehrlichial species. ELISA and Western blot analysis confirmed that the E. chaffeensis rMAP2 failed to serologically differentiate between E. canis and E. chaffeensis infections.

University of Florida College of Veterinary Medicine Journal Series number 631.

Present address: Department of Oral Biology, Center for Molecular Microbiology, University of Florida, Gainesville, FL 32610-0424.