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Journal of Clinical Microbiology, March 2005, p. 1309-1317, Vol. 43, No. 3
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.3.1309-1317.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Sequence Analysis of the msp4 Gene of Anaplasma phagocytophilum Strains

José de la Fuente,1,2* Robert F. Massung,3 Susan J. Wong,4 Frederick K. Chu,5 Hans Lutz,6 Marina Meli,6 Friederike D. von Loewenich,7 Anna Grzeszczuk,8 Alessandra Torina,9 Santo Caracappa,9 Atilio J. Mangold,10 Victoria Naranjo,2 Snorre Stuen,11 and Katherine M. Kocan1

Department of Veterinary Pathobiology, College of Veterinary Medicine, Oklahoma State University, Stillwater, Oklahoma,1 Division of Viral and Rickettsial Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia,3 Diagnostic Immunology Laboratory, Wadsworth Center,4 Wadsworth Center for Laboratories and Research, New York State Department of Health, Albany, New York,5 Instituto de Investigación en Recursos Cinegéticos IREC (CSIC-UCLM-JCCM), Ciudad Real, Spain,2 Clinical Laboratory, Faculty of Veterinary Medicine, University of Zurich, Zurich, Switzerland,6 Institute of Medical Microbiology and Hygiene, Department of Medical Microbiology and Hygiene, University Clinic of Freiburg, Freiburg, Germany,7 Department of Infectious Diseases, Medical University of Bialystok, Bialystok, Poland,8 Istituto Zooprofilattico Sperimentale della Sicilia, Palermo, Italy,9 Instituto Nacional de Tecnología Agropecuaria, Estación Experimental Agropecuaria Rafaela, Rafaela, Santa Fe, Argentina,10 Department of Production Animal Clinical Sciences, Norwegian School of Veterinary Science, Sandnes, Norway,11

Received 22 January 2004/ Returned for modification 1 July 2004/ Accepted 7 November 2004

The causative agent of human granulocytic ehrlichiosis was recently reclassified as Anaplasma phagocytophilum, unifying previously described bacteria that cause disease in humans, horses, dogs, and ruminants. For the characterization of genetic heterogeneity in this species, the homologue of Anaplasma marginale major surface protein 4 gene (msp4) was identified, and the coding region was PCR amplified and sequenced from a variety of sources, including 50 samples from the United States, Germany, Poland, Norway, Italy, and Switzerland and 4 samples of A. phagocytophilum-like organisms obtained from white-tailed deer in the United States. Sequence variation between strains of A. phagocytophilum (90 to 100% identity at the nucleotide level and 92 to 100% similarity at the protein level) was higher than in A. marginale. Phylogenetic analyses of msp4 sequences did not provide phylogeographic information but did differentiate strains of A. phagocytophilum obtained from ruminants from those obtained from humans, dogs, and horses. The sequence analysis of the recently discovered A. phagocytophilum msp2 gene corroborated these results. The results reported here suggest that although A. phagocytophilum-like organisms from white-tailed deer may be closely related to A. phagocytophilum, they could be more diverse. These results suggest that A. phagocytophilum strains from ruminants could share some common characteristics, including reservoirs and pathogenicity, which may be different from strains that infect humans.

* Corresponding author. Mailing address: Department of Veterinary Pathobiology, College of Veterinary Medicine, Oklahoma State University, Stillwater, OK 74078. Phone: (405) 744-0372. Fax: (405) 744-5275. E-mail: jose_delafuente{at}yahoo.com.

Journal of Clinical Microbiology, March 2005, p. 1309-1317, Vol. 43, No. 3
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.3.1309-1317.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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