Parasitology Division, National Veterinary Research Institute PMB 01, Vom, Plateau State, Nigeria.
Res Vet Sci. 2013 Feb;94(1):27-32. doi: 10.1016/j.rvsc.2012.07.031. Epub 2012 Aug 24.
The present study aimed to detect the presence of Ehrlichia canis in naturally infected dogs in Nigeria, using a combination of PCR and sequence analysis of the 16S rRNA gene and two genes encoding the tandem repeat-containing proteins (TRPs), TRP19 and TRP36. Out of a total of 100 blood samples collected from domestic dogs presented to veterinary hospitals in Jos, the capital city of Plateau State of Nigeria, 11 were positive in nested PCR for E. canis. Sequencing results for these amplicons showed that all of the 16S rDNA sequences (1623 bp) or the TRP19 coding sequences (414 bp) were identical to each other and had very high similarities (99.3-100%) with those from other E. canis strains accessible in GenBank. The TRP36 gene sequences derived from the 11 Nigerian isolates were identical to each other except for the number of the 27-bp repeat unit in a tandem repeat region, which was found to be 8, 12 or 18. Without considering the number of tandem repeats, these sequences had 100% identity to that of the reported Cameroon 71 isolate, but distinctly differed from those obtained from other geographically distant E. canis strains previously published. A phylogenetic tree of E. canis based on the TRP36 amino acid sequences showed that the Nigerian isolates and the Cameroon 71 isolate fell into a separate clade, indicating that they may share a common ancestor. Overall, this study not only provides the first molecular evidence of E. canis infections in dogs from Nigeria but also highlights the value of the TRP36 gene as a tool to classify E. canis isolates and to elucidate their phylogeographic relationships.
本研究旨在使用聚合酶链反应(PCR)结合 16S rRNA 基因和两个编码串联重复蛋白(TRP)的基因(TRP19 和 TRP36)的序列分析,检测尼日利亚自然感染犬中的埃立克体属犬种。从尼日利亚高原州首府乔斯的兽医医院采集的 100 份犬血液样本中,有 11 份经嵌套 PCR 检测为犬埃立克体属阳性。这些扩增子的测序结果表明,所有 16S rDNA 序列(1623bp)或 TRP19 编码序列(414bp)彼此之间完全相同,与 GenBank 中可获得的其他犬埃立克体属菌株具有非常高的相似性(99.3-100%)。从 11 株尼日利亚分离株中获得的 TRP36 基因序列彼此完全相同,但在串联重复区的 27-bp 重复单元数量上存在差异,分别为 8、12 或 18。不考虑串联重复数,这些序列与报道的喀麦隆 71 分离株完全相同,但与之前发表的其他地理上遥远的犬埃立克体属菌株明显不同。基于 TRP36 氨基酸序列的犬埃立克体属系统发育树显示,尼日利亚分离株和喀麦隆 71 分离株属于一个单独的分支,表明它们可能有共同的祖先。总的来说,本研究不仅为尼日利亚犬感染埃立克体属提供了首个分子证据,还突出了 TRP36 基因作为分类犬埃立克体属分离株和阐明其系统地理关系的工具的价值。