Dual Presence of Anaplasma phagocytophilum and Its Closely Related Anaplasma sp. in Ixodid Ticks in Hokkaido, Japan, and Their Specific Molecular Detection

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  • YBAÑEZ Adrian Patalinghug
    Department of Veterinary Clinical Science, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080–8555, Japan United Graduate School of Veterinary Sciences, Gifu University, Gifu 501–1193, Japan
  • MATSUMOTO Kotaro
    Department of Veterinary Clinical Science, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080–8555, Japan
  • KISHIMOTO Toshio
    Okayama Prefectural Institute for Environmental Science and Public Health, Okayama 701–0298, Japan
  • YOKOYAMA Naoaki
    National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080–8555, Japan
  • INOKUMA Hisashi
    Department of Veterinary Clinical Science, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080–8555, Japan United Graduate School of Veterinary Sciences, Gifu University, Gifu 501–1193, Japan

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  • Dual Presence of <i>Anaplasma phagocytophilum</i> and Its Closely Related <i>Anaplasma</i> sp. in Ixodid Ticks in Hokkaido, Japan, and Their Specific Molecular Detection

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Anaplasma phagocytophilum causes human granulocytic anaplasmosis (HGA) and tick-borne fever in ruminants. A closely related and potentially novel Anaplasma sp. in Japan was recently characterized. The aims of the study were to provide molecular evidence for the presence of these 2 species in Japan, and to develop a reliable PCR method based on the nucleotide differences within the citrate synthase (gltA) gene. DNA samples from 182 ixodid ticks (134 Ixodes persulcatus, 35 Haemaphysalis douglasii and 13 I. ovatus) collected from 2 sites in Hokkaido, Japan, were screened for A. phagocytophilum and its closely related Anaplasma sp. (herein designated as Anaplasma sp. Japan) using 16S rRNA PCR, revealing a combined prevalence rate of 27.5% (50 samples). The positive samples were then used to evaluate a newly developed gltA-based nested PCR method. Selected positive samples were further characterized using the groEL gene for confirmation and phylogenetic analyses. Two groups of sequence results were obtained: those that had closer identities with (1) A. phagocytophilum (99.5–99.6% for 16S rRNA, 97.5% for gltA and 98.4% for groEL), and those that had closer identities with (2) Anaplasma sp. closely related to A. phagocytophilum in Japan (99.3% for 16S rRNA, 96.4–98.7% for gltA and 97.5–97.9% for groEL). The present study confirmed the distinct presence of A. phagocytophilum and its closely related Anaplasma sp. in Japan, and developed a new PCR detection method based on gltA that can distinguish the 2 organisms.

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