title: A new multiple-locus variable-number tandem repeat analysis reveals different clusters for Anaplasma phagocytophilum circulating in domestic and wild ruminants.
abstract: BACKGROUND
  Anaplasma phagocytophilum is a tick-borne intragranulocytic alpha-proteobacterium. It is the causative agent of tick-borne fever in ruminants, and of human granulocytic anaplasmosis in humans, two diseases which are becoming increasingly recognized in Europe and the USA. However, while several molecular typing tools have been developed over the last years, few of them are appropriate for in-depth exploration of the epidemiological cycle of this bacterium. Therefore we have developed a Multiple-Locus Variable number tandem repeat (VNTR) Analysis typing technique for A. phagocytophilum.
  
  
  METHODS
  Five VNTRs were selected based on the HZ human-derived strain genome, and were tested on the Webster human-derived strain and on 123 DNA samples: 67 from cattle, 7 from sheep, 15 from roe deer, 4 from red deer, 1 from a reindeer, 2 from horses, 1 from a dog, and 26 from ticks.
  
  
  RESULTS
  From these samples, we obtained 84 different profiles, with a diversity index of 0.96 (0.99 for vertebrate samples, i.e. without tick samples). Our technique confirmed that A. phagocytophilum from roe deer or domestic ruminants belong to two different clusters, while A. phagocytophilum from red deer and domestic ruminants locate within the same cluster, questioning the respective roles of roe vs red deer as reservoir hosts for domestic ruminant strains in Europe. As expected, greater diversity was obtained between rather than within cattle herds.
  
  
  CONCLUSIONS
  Our technique has great potential to provide detailed information on A. phagocytophilum isolates, improving both epidemiological and phylogenic investigations, thereby helping in the development of relevant prevention and control measures.
authors.forename: Thibaud
authors.initials: T
authors.lastname: Dugat
authors.forename: Amélie
authors.initials: A
authors.lastname: Chastagner
authors.forename: Anne-Claire
authors.initials: AC
authors.lastname: Lagrée
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Petit
authors.forename: Benoît
authors.initials: B
authors.lastname: Durand
authors.forename: Simon
authors.initials: S
authors.lastname: Thierry
authors.forename: Fabien
authors.initials: F
authors.lastname: Corbière
authors.forename: Hélène
authors.initials: H
authors.lastname: Verheyden
authors.forename: Luc
authors.initials: L
authors.lastname: Chabanne
authors.forename: Xavier
authors.initials: X
authors.lastname: Bailly
authors.forename: Agnès
authors.initials: A
authors.lastname: Leblond
authors.forename: Gwenaël
authors.initials: G
authors.lastname: Vourc'h
authors.forename: Henri-Jean
authors.initials: HJ
authors.lastname: Boulouis
authors.affiliation: Université Paris-Est, Ecole Nationale Vétérinaire d'Alfort, UMR BIPAR ENVA Anses UPEC USC INRA, Maisons-Alfort, France. r.maillard@envt.fr.
authors.forename: Renaud
authors.initials: R
authors.lastname: Maillard
authors.forename: Nadia
authors.initials: N
authors.lastname: Haddad
mesh_terms: D041081:Anaplasma phagocytophilum / Q000235:genetics* / Q000302:isolation & purification; D000818:Animals; D000829:Animals, Domestic; D000835:Animals, Wild; D015373:Bacterial Typing Techniques; D001483:Base Sequence; D016000:Cluster Analysis; D004197:Disease Reservoirs / Q000382:microbiology; D016873:Ehrlichiosis / Q000382:microbiology / Q000662:veterinary*; D005602:France; D056426:Genetic Loci / Q000235:genetics; D014644:Genetic Variation; D005843:Geography; D006801:Humans; D018598:Minisatellite Repeats / Q000235:genetics*; D008969:Molecular Sequence Data; D010802:Phylogeny; D012418:Ruminants; D017422:Sequence Analysis, DNA; D020080:Tandem Repeat Sequences / Q000235:genetics*; D017282:Tick-Borne Diseases / Q000382:microbiology / Q000662:veterinary*