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对来自加拿大不列颠哥伦比亚省的自由放养骡鹿和白尾鹿进行分子研究,以寻找 Anaplasma spp. 和 Ehrlichia spp. 的证据。

Molecular study of free-ranging mule deer and white-tailed deer from British Columbia, Canada, for evidence of Anaplasma spp. and Ehrlichia spp.

机构信息

Canadian Food Inspection Agency, Centre for Food-Borne and Animal Parasitology, Saskatoon, SK, Canada.

出版信息

Transbound Emerg Dis. 2012 Jun;59(3):233-43. doi: 10.1111/j.1865-1682.2011.01259.x. Epub 2011 Sep 20.

Abstract

Twenty-three free-ranging white-tailed deer (WTD; Odocoileus virginianus) and six mule deer (MD; Odocoileus hemionus) from south-central British Columbia, Canada, were tested for Anaplasma marginale by msp5 gene-specific PCR and Ehrlichia spp. by 16S rRNA or citrate synthase (gltA) gene-specific PCR, as well as by PCR with universal 16S rRNA primers detecting a wide range of bacteria. No deer tested positive for A. marginale. Amplification with universal 16S rRNA primers followed by sequencing of cloned fragments detected an Anaplasma sp. in one of 23 (4.3%) WTD and six of six (100%) MD and Bartonella sp. in four of 23 (17.4%) WTD. The Anaplasma sp. was genetically distinct from A. marginale and all other recognized members of the genus. Four of six (66.7%) MD and 0 of 23 (0%) WTD were Ehrlichia positive by PCR with primers for 16S rRNA and gltA genes. The sequences of gltA PCR fragments were identical to each other and to the respective region of the gltA gene of an Ehrlichia sp. which we detected previously in naturally infected cattle from the same area, suggesting the possibility of biological transmission of this rickettsia between cattle and wild cervids. Antibodies reactive with the MSP5 protein of A. marginale were detected using a competitive enzyme-linked immunosorbent assay in two of six (33.3%) MD, but not in WTD. The two seropositive MD were PCR positive for both the Anaplasma sp. and Ehrlichia sp. detected in this study, suggesting a reaction of antibodies against one or both of these rickettsias with the MSP5 antigen.

摘要

23 头来自加拿大不列颠哥伦比亚省中南部的自由放养白尾鹿(Odocoileus virginianus)和 6 头骡鹿(Odocoileus hemionus)接受了针对边缘无浆体(Anaplasma marginale)的 msp5 基因特异性 PCR 检测和埃立克体属(Ehrlichia spp.)的 16S rRNA 或柠檬酸合酶(gltA)基因特异性 PCR 检测,以及使用通用 16S rRNA 引物的 PCR 检测,该引物可检测广泛的细菌。没有鹿对边缘无浆体呈阳性。使用通用 16S rRNA 引物进行扩增,然后对克隆片段进行测序,在 23 头白尾鹿中的 1 头(4.3%)和 6 头骡鹿中的 6 头(100%)中检测到一种属于无形体属(Anaplasma sp.)的细菌,在 23 头白尾鹿中的 4 头(17.4%)中检测到一种巴尔通体属(Bartonella sp.)的细菌。该无形体属与边缘无浆体和所有其他公认的属成员在基因上是不同的。6 头骡鹿中的 4 头(66.7%)和 23 头白尾鹿中的 0 头(0%)通过针对 16S rRNA 和 gltA 基因的引物进行 PCR 检测为埃立克体属阳性。gltA PCR 片段的序列彼此相同,并且与我们之前在同一地区自然感染的牛中检测到的埃立克体属的 gltA 基因的相应区域相同,这表明这种立克次体在牛和野生鹿科动物之间存在生物传播的可能性。使用竞争酶联免疫吸附试验(ELISA)检测到两种骡鹿(33.3%)的抗边缘无浆体 MSP5 蛋白的抗体,但在白尾鹿中未检测到。这两例血清阳性的骡鹿对本研究中检测到的无形体属和埃立克体属均为 PCR 阳性,这表明针对这些立克次体之一或两者的抗体与 MSP5 抗原发生反应。

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