Specific Molecular Detection and Characterization of Anaplasma marginale in Mongolian Cattle
Specific Molecular Detection and Characterization of Anaplasma marginale in Mongolian Cattle
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DOI：
10.1292/jvms.12-0361
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发表时间：
2013-04-01
影响因子：
1.2
通讯作者：
Inokuma, Hisashi
中科院分区：
文献类型：
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作者：
Ybanez, Adrian Patalinghug;Sivakumar, Thillaiampalam;Inokuma, Hisashi
边缘无形体是牛无形体病的病原体。本研究旨在从分子水平检测和表征A。在蒙古牛种群中流行的边缘牛。建立了一种基于Msp 5基因的高特异性、高灵敏度的巢式PCR（nPCR）方法。边缘化（Msp 5 nPCR）。该方法检测到A.从不同国家获得的阳性DNA样品中观察到边缘扩增子，而从测试的几种其他牛血液病原体的DNA样品中未观察到扩增子。Msp 5 nPCR的检测限为2拷贝/μ l。2010年，对300头蒙古牛的田间血液DNA样品进行了检测。结果显示，患病率为8.7%（26/300）。另一方面，与A.绵羊（同源性为95.0%）的基因进行了检测。基于Msp 5、groEL和16 SrRNA基因的系统发育分析表明，A.蒙古的边缘分离物与分布于其他国家的分离物没有区别。本研究成功地建立了一种新的检测A. marginate等首次报道了A. marginate和一个与A.蒙古牛种群中的绵羊。
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. marginate (Msp5 nPCR). The method detected A. marginate 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/mu l. 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. marginate 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. marginate, and reported the first molecular detection and characterization of A. marginate and an Anaplasma sp. closely related to A. ovis in Mongolian cattle populations.