title: Multilocus Sequence Analysis of Clinical "Candidatus Neoehrlichia mikurensis" Strains from Europe.
abstract: "Candidatus Neoehrlichia mikurensis" is the tick-borne agent of neoehrlichiosis, an infectious disease that primarily affects immunocompromised patients. So far, the genetic variability of "Ca. Neoehrlichia" has been studied only by comparing 16S rRNA genes and groEL operon sequences. We describe the development and use of a multilocus sequence analysis (MLSA) protocol to characterize the genetic diversity of clinical "Ca. Neoehrlichia" strains in Europe and their relatedness to other species within the Anaplasmataceae family. Six genes were selected: ftsZ, clpB, gatB, lipA, groEL, and 16S rRNA. Each MLSA locus was amplified by real-time PCR, and the PCR products were sequenced. Phylogenetic trees of MLSA locus relatedness were constructed from aligned sequences. Blood samples from 12 patients with confirmed "Ca. Neoehrlichia" infection from Sweden (n = 9), the Czech Republic (n = 2), and Germany (n = 1) were analyzed with the MLSA protocol. Three of the Swedish strains exhibited identical lipA sequences, while the lipA sequences of the strains from the other nine patients were identical to each other. One of the Czech strains had one differing nucleotide in the clpB sequence from the sequences of the other 11 strains. All 12 strains had identical sequences for the genes 16S rRNA, ftsZ, gatB, and groEL. According to the MLSA, among the Anaplasmataceae, "Ca. Neoehrlichia" is most closely related to Ehrlichia ruminantium, less so to Anaplasma phagocytophilum, and least to Wolbachia endosymbionts. To conclude, three sequence types of infectious "Ca. Neoehrlichia" were identified: one in the west of Sweden, one in the Czech Republic, and one spread throughout Europe.
authors.affiliation: Department of Clinical Microbiology, Sahlgrenska University Hospital, and Department of Infectious Diseases, Sahlgrenska Academy at the University of Gothenburg, Gothenburg, Sweden.
authors.forename: Anna
authors.initials: A
authors.lastname: Grankvist
authors.affiliation: Department of Clinical Microbiology, Sahlgrenska University Hospital, and Department of Infectious Diseases, Sahlgrenska Academy at the University of Gothenburg, Gothenburg, Sweden.
authors.forename: Edward R B
authors.initials: ER
authors.lastname: Moore
authors.affiliation: Department of Clinical Microbiology, Sahlgrenska University Hospital, and Department of Infectious Diseases, Sahlgrenska Academy at the University of Gothenburg, Gothenburg, Sweden.
authors.forename: Liselott
authors.initials: L
authors.lastname: Svensson Stadler
authors.affiliation: Laboratory for Molecular Diagnostics, CHAMBON Laboratories, Prague, Czech Republic.
authors.forename: Sona
authors.initials: S
authors.lastname: Pekova
authors.affiliation: Mikrobiologisches Institut, Klinische Mikrobiologie, Immunologie und Hygiene, Friedrich Alexander Universität (FAU) Erlangen-Nürnberg, and Universitätsklinikum Erlangen, Erlangen, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Bogdan
authors.affiliation: Mikrobiologisches Institut, Klinische Mikrobiologie, Immunologie und Hygiene, Friedrich Alexander Universität (FAU) Erlangen-Nürnberg, and Universitätsklinikum Erlangen, Erlangen, Germany.
authors.forename: Walter
authors.initials: W
authors.lastname: Geißdörfer
authors.affiliation: Department of Medicine, Kungälv Hospital, Kungälv, Sweden.
authors.forename: Jenny
authors.initials: J
authors.lastname: Grip-Lindén
authors.affiliation: Department of Infectious Diseases, Skaraborg Hospital, Skövde, Sweden.
authors.forename: Kenny
authors.initials: K
authors.lastname: Brandström
authors.affiliation: Department of Experimental Medical Science, Section of Immunology, and Department of Clinical Science, Section of Medicine, Lund University, Lund, Sweden.
authors.forename: Jan
authors.initials: J
authors.lastname: Marsal
authors.affiliation: Department of Clinical Sciences, Section of Rheumatology, Lund University, Lund, Sweden.
authors.forename: Kristofer
authors.initials: K
authors.lastname: Andréasson
authors.affiliation: Department of Hematology and Coagulation, Sahlgrenska University Hospital, Göteborg, Sweden.
authors.forename: Catharina
authors.initials: C
authors.lastname: Lewerin
authors.affiliation: Department of Clinical Microbiology, Sahlgrenska University Hospital, and Department of Infectious Diseases, Sahlgrenska Academy at the University of Gothenburg, Gothenburg, Sweden.
authors.forename: Christina
authors.initials: C
authors.lastname: Welinder-Olsson
authors.affiliation: Department of Clinical Microbiology, Sahlgrenska University Hospital, and Department of Infectious Diseases, Sahlgrenska Academy at the University of Gothenburg, Gothenburg, Sweden christine.wenneras@microbio.gu.se.
authors.forename: Christine
authors.initials: C
authors.lastname: Wennerås
mesh_terms: D000368:Aged; D000710:Anaplasmataceae / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D000711:Anaplasmataceae Infections / Q000453:epidemiology / Q000382:microbiology*; D016000:Cluster Analysis; D018153:Czech Republic / Q000453:epidemiology; D005260:Female; D020043:Genes, Essential; D014644:Genetic Variation; D005838:Genotype; D005858:Germany / Q000453:epidemiology; D006801:Humans; D008297:Male; D008875:Middle Aged; D017720:Molecular Epidemiology / Q000379:methods; D058885:Multilocus Sequence Typing / Q000379:methods*; D010802:Phylogeny; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D013548:Sweden / Q000453:epidemiology
chemical_list: D012336:RNA, Ribosomal, 16S