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Journal of Clinical Microbiology, June 2007, p. 1771-1776, Vol. 45, No. 6
0095-1137/07/$08.00+0     doi:10.1128/JCM.00365-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

High-Resolution Genetic Fingerprinting of European Strains of Anaplasma phagocytophilum by Use of Multilocus Variable-Number Tandem-Repeat Analysis

Infectious Disease Ecology Group, Faculty of Veterinary Science, University of Liverpool, Leahurst, Chester High Rd., Neston, Cheshire CH64 4EF, United Kingdom,¹ Aberdeen Population Ecology Unit, School of Biological Sciences, University of Aberdeen, Tillydrone Avenue, Aberdeen AB24 2TZ, United Kingdom,² Centre for Infectious Disease Prevention and Control, Public Health Agency of Canada, Universite de Montreal, Montreal, Quebec J2S 7C6, Canada,³ Department of Infectious Diseases, University Medical Center Ljubljana, University of Ljubljana, Ljubljana, Slovenia,⁴ School of Clinical Veterinary Science, University of Bristol, Langford House, Langford, Bristol BS40 5DU, United Kingdom⁵

Received 15 February 2007/ Returned for modification 27 March 2007/ Accepted 9 April 2007

Anaplasma phagocytophilum is a widely distributed tick-borne pathogen of humans, livestock, and companion animals. We used in silico methods to identify 10 variable-number tandem-repeat (VNTR) loci within the genome sequence of the A. phagocytophilum HZ strain and used these data to develop a multilocus VNTR-based typing scheme for the species. Having confirmed the stability of four of the loci in replicates of the A. phagocytophilum strain that had been subjected to different numbers of passages through cell cocultures in vitro, we then used this typing scheme to discriminate between 20 A. phagocytophilum strains of diverse geographical and host provenances. Extensive diversity was found at each of the four loci studied, with total allele numbers ranging from 13 to 18 and Hunter-Gaston discriminatory index values ranging from 0.93 to 0.99. Only 2 of the 20 strains examined shared alleles at all four loci. The discriminatory power of VNTR analysis was found to be greater than that of either partial msp4 or 16S rRNA gene sequence comparison. The extremely high sensitivity of this novel approach to the genetic fingerprinting of A. phagocytophilum strains should serve well in molecular epidemiological studies of infection transmission, particularly when fine-scale strain delineation is required.

Published ahead of print on 18 April 2007.

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