title: Specific molecular detection and characterization of Anaplasma marginale in Mongolian cattle.
abstract: Anaplasma marginale is an etiologic agent of bovine anaplasmosis. This study aimed to molecularly detect and characterize A. marginale that is prevalent in Mongolian cattle populations. A highly specific and sensitive nested PCR (nPCR) method based on the Msp5 gene was developed to detect A. marginale (Msp5 nPCR). The method detected A. marginale from the positive DNA samples obtained from different countries, while no amplicons were observed from DNA samples of several other bovine blood pathogens tested. The detection limit of Msp5 nPCR was determined to be 2 copies/μl. The method was tested against field blood DNA samples prepared from 300 Mongolian cattle in 2010. Results indicated a prevalence rate of 8.7% (26 of 300). On the other hand, partial DNA fragments of an Anaplasma sp. closely related to A. ovis (with 95.0% identity) were detected using a different nPCR method based on groEL gene. The phylogenetic analyses based on the Msp5, groEL and 16S rRNA genes demonstrated that A. marginale isolates in Mongolia were not divergent from the isolates distributed in other countries. The present study successfully established a new nPCR assay that can detect A. marginale, and reported the first molecular detection and characterization of A. marginale and an Anaplasma sp. closely related to A. ovis in Mongolian cattle populations.
authors.affiliation: Department of Veterinary Clinical Sciences, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido, Japan.
authors.forename: Adrian Patalinghug
authors.initials: AP
authors.lastname: Ybañez
authors.forename: Thillaiampalam
authors.initials: T
authors.lastname: Sivakumar
authors.forename: Badgar
authors.initials: B
authors.lastname: Battsetseg
authors.forename: Banzragch
authors.initials: B
authors.lastname: Battur
authors.forename: Khukhuu
authors.initials: K
authors.lastname: Altangerel
authors.forename: Kotaro
authors.initials: K
authors.lastname: Matsumoto
authors.forename: Naoaki
authors.initials: N
authors.lastname: Yokoyama
authors.forename: Hisashi
authors.initials: H
authors.lastname: Inokuma
mesh_terms: D042322:Anaplasma marginale / Q000235:genetics / Q000302:isolation & purification*; D000712:Anaplasmosis / Q000453:epidemiology / Q000382:microbiology*; D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000235:genetics; D001483:Base Sequence; D002417:Cattle; D002418:Cattle Diseases / Q000453:epidemiology / Q000382:microbiology*; D018834:Chaperonin 60 / Q000737:chemistry / Q000235:genetics; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D008969:Molecular Sequence Data; D008986:Mongolia / Q000453:epidemiology; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000662:veterinary; D015995:Prevalence; D012336:RNA, Ribosomal, 16S / Q000737:chemistry / Q000235:genetics; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
chemical_list: D001425:Bacterial Outer Membrane Proteins; D018834:Chaperonin 60; D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S; C111578:major surface protein 5, Anaplasma