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Journal of Clinical Microbiology, October 2000, p. 3705-3709, Vol. 38, No. 10
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Expression of a Gene Encoding the Major Antigenic Protein 2 Homolog of Ehrlichia chaffeensis and Potential Application for Serodiagnosis

A. Rick Alleman,1,* Anthony F. Barbet,2 Michael V. Bowie,2 Heather L. Sorenson,1 Susan J. Wong,3 and Myriam Bélanger2

Departments of Physiological Sciences1 and Pathobiology,2 College of Veterinary Medicine, University of Florida, Gainesville, Florida 32610, and Wadsworth Center, New York State Department of Health, Albany, New York 122013

Received 8 February 2000/Returned for modification 3 May 2000/Accepted 10 July 2000

The major antigenic protein 2 (MAP2) homolog of Ehrlichia chaffeensis 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 human 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. However, immune sera failed to react with the recombinant on immunoblots when the antigen was denatured by heat or reduced using beta -mercaptoethanol. The recombinant MAP2 (rMAP2) was used in an ELISA format with 60 blinded serum samples. Twenty of the serum samples were previously demonstrated to contain antibodies reactive with E. chaffeensis by indirect immunofluorescence assays (IFAs). The remaining 40 samples were seronegative. All samples negative by IFA were also found to be negative for antibodies against the rMAP2 of E. chaffeensis by using the ELISA. Only 1 of 20 IFA-positive samples tested negative in the rMAP2 ELISA. There was 100% agreement using IFA-negative samples and 95% agreement using IFA-positive samples, resulting in a 97.5% overall agreement between the two assays. These data suggest that the rMAP2 homolog of E. chaffeensis may have potential as a test antigen for the serodiagnosis of human monocytic ehrlichiosis. To our knowledge, this recombinant is unique because it is thus far the only E. chaffeensis recombinant antigen that has been shown to work in an ELISA format.

* 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, October 2000, p. 3705-3709, Vol. 38, No. 10
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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