title: Specific molecular detection of Anaplasma sp. closely related to Anaplasma phagocytophilum in ixodid ticks and cattle in a pastureland in Hokkaido, Japan.
abstract: Recent molecular analyses of the Anaplasma sp. closely related to Anaplasma phagocytophilum (previously believed to be A. phagocytophilum) in Japan have clarified its distinct phylogenetic position. PCR methods relying on 16S rRNA- and P44/MSP2-based primers designed to detect this species have low sensitivity and specificity. In this study, a highly sensitive and specific nested PCR method using newly designed primers based on heat-shock operon gene (groEL) was developed to detect this species. The method was later used in an epidemiological study testing DNA samples from 85 Ixodid ticks (collected by flagging) and 50 cattle from the same pastureland in Nakaosobetsu, Hokkaido, Japan. Results revealed prevalence rates of 2.4% (2 of 85) in ticks and 2% (1 of 50) in cattle. The present study also reported the first molecular detection of the Anaplasma sp. closely related to A. phagocytophilum in Japan in H. douglasii, and established a new reliable PCR method that detects this Anaplasma sp. closely related to A. phagocytophilum in Japan.
authors.affiliation: Department of Veterinary Clinical Sciences, Obihiro University of Agriculture and Veterinary Medicine, Inada Cho, Hokkaido, Japan.
authors.forename: Adrian P
authors.initials: AP
authors.lastname: Ybañez
authors.forename: Michihito
authors.initials: M
authors.lastname: Tagawa
authors.forename: Kotaro
authors.initials: K
authors.lastname: Matsumoto
authors.forename: Toshio
authors.initials: T
authors.lastname: Kishimoto
authors.forename: Naoaki
authors.initials: N
authors.lastname: Yokoyama
authors.forename: Hisashi
authors.initials: H
authors.lastname: Inokuma
mesh_terms: D000709:Anaplasma / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D041081:Anaplasma phagocytophilum / Q000235:genetics / Q000302:isolation & purification; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D002417:Cattle; D018834:Chaperonin 60 / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D016873:Ehrlichiosis / Q000453:epidemiology / Q000382:microbiology*; D018884:Ixodes / Q000382:microbiology*; D007564:Japan / Q000453:epidemiology; D008297:Male; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D015995:Prevalence; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013045:Species Specificity
chemical_list: D001426:Bacterial Proteins; D018834:Chaperonin 60; D017931:DNA Primers; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S