Kimita Gathii, Mutai Beth, Nyanjom Steven Ger, Wamunyokoli Fred, Waitumbi John
Walter Reed Project/ Kenya Medical Research Institute, Kisumu, Kenya.
Department of Biochemistry, Jomo Kenyatta University of Agriculture and Technology, Juja, Kenya.
PLoS Negl Trop Dis. 2016 Jul 7;10(7):e0004788. doi: 10.1371/journal.pntd.0004788. eCollection 2016 Jul.
Rickettsia africae, the etiological agent of African tick bite fever, is widely distributed in sub-Saharan Africa. Contrary to reports of its homogeneity, a localized study in Asembo, Kenya recently reported high genetic diversity. The present study aims to elucidate the extent of this heterogeneity by examining archived Rickettsia africae DNA samples collected from different eco-regions of Kenya.
To evaluate their phylogenetic relationships, archived genomic DNA obtained from 57 ticks a priori identified to contain R. africae by comparison to ompA, ompB and gltA genes was used to amplify five rickettsial genes i.e. gltA, ompA, ompB, 17kDa and sca4. The resulting amplicons were sequenced. Translated amino acid alignments were used to guide the nucleotide alignments. Single gene and concatenated alignments were used to infer phylogenetic relationships.
Out of the 57 DNA samples, three were determined to be R. aeschlimanii and not R. africae. One sample turned out to be a novel rickettsiae and an interim name of "Candidatus Rickettsia moyalensis" is proposed. The bonafide R. africae formed two distinct clades. Clade I contained 9% of the samples and branched with the validated R. africae str ESF-5, while clade II (two samples) formed a distinct sub-lineage.
This data supports the use of multiple genes for phylogenetic inferences. It is determined that, despite its recent emergence, the R. africae lineage is diverse. This data also provides evidence of a novel Rickettsia species, Candidatus Rickettsia moyalensis.
非洲立克次体是非洲蜱咬热的病原体,广泛分布于撒哈拉以南非洲地区。与关于其同质性的报道相反,肯尼亚阿森博的一项局部研究最近报告了其高度的遗传多样性。本研究旨在通过检查从肯尼亚不同生态区域收集的存档非洲立克次体DNA样本,阐明这种异质性的程度。
为了评估它们的系统发育关系,使用通过与ompA、ompB和gltA基因比较预先鉴定为含有非洲立克次体的57只蜱虫的存档基因组DNA,扩增五个立克次体基因,即gltA、ompA、ompB、17kDa和sca4。对产生的扩增子进行测序。使用翻译后的氨基酸比对来指导核苷酸比对。单基因比对和串联比对用于推断系统发育关系。
在57个DNA样本中,有三个被确定为埃塞利曼立克次体而非非洲立克次体。一个样本被证明是一种新型立克次体,并提出了一个临时名称“暂定莫亚勒立克次体”。真正的非洲立克次体形成了两个不同的进化枝。进化枝I包含9%的样本,并与经过验证的非洲立克次体菌株ESF-5分支,而进化枝II(两个样本)形成了一个独特的亚谱系。
这些数据支持使用多个基因进行系统发育推断。已确定,尽管非洲立克次体谱系最近才出现,但它是多样的。这些数据还提供了一种新型立克次体物种——暂定莫亚勒立克次体的证据。