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Journal of Clinical Microbiology, July 2001, p. 2494-2499, Vol. 39, No. 7
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.7.2494-2499.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Recombinant Major Antigenic Protein 2 of Ehrlichia canis: a Potential Diagnostic Tool

A. Rick Alleman,1,* Leo J. McSherry,1 Anthony F. Barbet,2 Edward B. Breitschwerdt,3 Heather L. Sorenson,1 Michael V. Bowie,2 and Myriam Bélanger2

Departments of Physiological Sciences1 and Pathobiology, 2 College of Veterinary Medicine, University of Florida, Gainesville, Florida 32610, and Department of Companion Animal and Special Species Medicine, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina 276063

Received 16 November 2000/Returned for modification 4 March 2001/Accepted 8 April 2001

The major antigenic protein 2 (MAP2) of Ehrlichia canis was cloned and expressed. The recombinant protein was characterized and tested in an enzyme-linked immunosorbent assay (ELISA) format for potential application in the serodiagnosis of canine monocytic ehrlichiosis. The recombinant protein, which contained a C-terminal polyhistidine tag, had a molecular mass of approximately 26 kDa. The antigen was clearly identified by Western immunoblotting using antihistidine antibody and immune serum from an experimentally infected dog. The recombinant MAP2 (rMAP2) was tested in an ELISA format using 141 serum samples from E. canis immunofluorescent antibody (IFA)-positive and IFA-negative dogs. Fifty-five of the serum samples were from dogs experimentally or naturally infected with E. canis and were previously demonstrated to contain antibodies reactive with E. canis by indirect immunofluorescence assays. The remaining 86 samples, 33 of which were from dogs infected with microorganisms other than E. canis, were seronegative. All of the samples from experimentally infected animals and 36 of the 37 samples from naturally infected animals were found to contain antibodies against rMAP2 of E. canis in the ELISA. Only 3 of 53 IFA-negative samples tested positive on the rMAP2 ELISA. There was 100% agreement among IFA-positive samples from experimentally infected animals, 97.3% agreement among IFA-positive samples from naturally infected animals, and 94.3% agreement among IFA-negative samples, resulting in a 97.2% overall agreement between the two assays. These data suggest that rMAP2 of E. canis could be used as a recombinant test antigen for the serodiagnosis of canine monocytic ehrlichiosis.

* Corresponding author. Mailing address: Department of Physiological Sciences, College of Veterinary Medicine, University of Florida, Box 100103C, Gainesville, FL 32610. Phone: (352) 392-4700, ext. 5858. Fax: (352) 392-1769. E-mail: ALLEMANR{at}MAIL.VETMED.UFL.EDU.

Journal of Clinical Microbiology, July 2001, p. 2494-2499, Vol. 39, No. 7
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.7.2494-2499.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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