Open AccessHigh-Resolution Genetic Fingerprinting of European Strains of Anaplasma phagocytophilum by Use of Multilocus Variable-Number Tandem-Repeat Analysis
Author(s) -
Kevin Bown,
Xavier Lambin,
Nicholas H. Ogden,
Miroslav Petrovec,
Susan E. Shaw,
Z. Woldehiwet,
Richard J. Birtles
Publication year - 2007
Publication title -
journal of clinical microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.349
H-Index - 255
eISSN - 1070-633X
pISSN - 0095-1137
DOI - 10.1128/jcm.00365-07
Subject(s) - biology , anaplasma phagocytophilum , genetics , variable number tandem repeat , tandem repeat , multilocus sequence typing , typing , multiple loci vntr analysis , anaplasma , allele , molecular epidemiology , genotype , tick , genome , gene , virology , borrelia burgdorferi , antibody
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 theA. 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 theA. 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 20A. 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 partialmsp4 or 16S rRNA gene sequence comparison. The extremely high sensitivity of this novel approach to the genetic fingerprinting ofA. phagocytophilum strains should serve well in molecular epidemiological studies of infection transmission, particularly when fine-scale strain delineation is required.