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Clinical and Vaccine Immunology, March 2007, p. 262-268, Vol. 14, No. 3
1071-412X/07/$08.00+0     doi:10.1128/CVI.00320-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Characterization of Anaplasma phagocytophilum Major Surface Protein 5 and the Extent of Its Cross-Reactivity with A. marginale{triangledown}

N. I. Strik,1 A. R. Alleman,1* A. F. Barbet,1 H. L. Sorenson,1 H. L. Wamsley,1 F. P. Gaschen,2,{dagger} N. Luckschander,2 S. Wong,3 F. Chu,3 J. E. Foley,4 A. Bjoersdorff,5 S. Stuen,6 and D. P. Knowles7

College of Veterinary Medicine, University of Florida, Gainesville, Florida,1 Vetsuisse Faculty, University of Bern, Bern, Switzerland,2 Wadsworth Center, New York State Department of Health, Albany, New York,3 Department of Medicine and Epidemiology, University of California, Davis, California,4 Department of Clinical Microbiology, Kalmar County Hospital, Kalmar, Sweden,5 Department of Sheep and Goat Research, Norwegian School of Veterinary Science, Sandnes, Norway,6 Animal Disease Research Unit, Agricultural Research Service, USDA, and Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, Washington7

Received 1 September 2006/ Returned for modification 16 October 2006/ Accepted 2 January 2007

Major surface protein 5 (Msp5) of Anaplasma marginale is highly conserved in the genus Anaplasma and the antigen used in a commercially available competitive enzyme-linked immunosorbent assay (cELISA) for serologic identification of cattle with anaplasmosis. This study analyzes the degrees of conservation of Msp5 among various isolates of Anaplasma phagocytophilum and the extent of serologic cross-reactivity between recombinant Msp5 (rMsp5) of Anaplasma marginale and A. phagocytophilum. The msp5 genes from various isolates of A. phagocytophilum were sequenced and compared. rMsp5 proteins of A. phagocytophilum and A. marginale were used separately in an indirect ELISA to detect cross-reactivity in serum samples from humans and dogs infected with A. phagocytophilum and cattle infected with A. marginale. Serum samples were also tested with a commercially available competitive ELISA that uses monoclonal antibody ANAF16C1. There were 100% sequence identities in the msp5 genes among all of the A. phagocytophilum isolates from the United States and a horse isolate from Sweden. Sheep isolates from Norway and dog isolates from Sweden were 99% identical to one another but differed in 17 base pairs from the United States isolates and the horse isolate. Serologic cross-reactivity was identified when serum samples from cattle infected with A. marginale were reacted with rMsp5 of A. phagocytophilum and when serum samples from humans and dogs infected with A. phagocytophilum were reacted with rMsp5 of A. marginale in an indirect-ELISA format. Serum samples from dogs or humans infected with A. phagocytophilum did not cross-react with rMsp5 of A. marginale when tested with the commercially available cELISA. These results suggest that rMsp5 of A. phagocytophilum is highly conserved among United States and European isolates and that serologic distinction between A. phagocytophilum and A. marginale infections cannot be accomplished if rMsp5 from either organism is used in an indirect ELISA.

* Corresponding author. Mailing address: College of Veterinary Medicine, University of Florida, 2015 S.W. 16th Ave., Box 100103, Gainesville, FL 32610. Phone: (352) 392-4700. Fax: (353) 392-2938. E-mail: AllemanR{at}mail.vetmed.ufl.edu.

{triangledown} Published ahead of print on 10 January 2007.

{dagger} Present address: School of Veterinary Medicine, Louisiana State University.

Clinical and Vaccine Immunology, March 2007, p. 262-268, Vol. 14, No. 3
1071-412X/07/$08.00+0     doi:10.1128/CVI.00320-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Ge, Y., Rikihisa, Y. (2007). Identification of Novel Surface Proteins of Anaplasma phagocytophilum by Affinity Purification and Proteomics. J. Bacteriol. 189: 7819-7828 [Abstract] [Full Text]  


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