Marosevic Durdica, Margos Gabriele, Wallich Reinhard, Wieser Andreas, Sing Andreas, Fingerle Volker
German National Reference Centre for Borrelia, Bavarian Health and Food Safety Authority (LGL), Oberschleißheim, Germany.
European Programme for Public Health Microbiology Training, European Centre of Disease Prevention and Control (ECDC), Stockholm, Sweden.
PLoS Negl Trop Dis. 2017 Sep 13;11(9):e0005865. doi: 10.1371/journal.pntd.0005865. eCollection 2017 Sep.
Borrelia recurrentis is the causative agent of louse-borne relapsing fever, endemic to the Horn of Africa. New attention was raised in Europe, with the highest number of cases (n = 45) reported among migrants in 2015 in Germany and sporadically from other European countries. So far only one genome was sequenced, hindering the development of specific molecular diagnostic and typing tools. Here we report on modified culture conditions for B. recurrentis and the intraspecies genome variability of six isolates isolated and cultured in different years in order to explore the possibility to identify new targets for typing and examine the molecular epidemiology of the pathogen.
METHODOLOGY/PRINCIPAL FINDINGS: Two historical isolates from Ethiopia and four isolates from migrants from Somalia (n = 3) and Ethiopia (n = 1) obtained in 2015 were cultured in MPK-medium supplemented with 50% foetal calf serum. Whole DNA was sequenced using Illumina MiSeq technology and analysed using the CLC Genomics Workbench and SPAdes de novo assembler. Compared to the reference B. recurrentis A1 29-38 SNPs were identified in the genome distributed on the chromosome and plasmids. In addition to that, plasmids of differing length, compared to the available reference genome were identified.
CONCLUSIONS/SIGNIFICANCE: The observed low genetic variability of B. recurrentis isolates is possibly due to the adaptation to a very conserved vector-host (louse-human) cycle, or influenced by the fastidious nature of the pathogen and their resistance to in vitro growth. Nevertheless, isolates obtained in 2015 were bearing the same chromosomal SNPs and could be distinguished from the historical isolates by means of whole genome sequencing, but not hitherto used typing methods. This is the first study examining the molecular epidemiology of B. recurrentis and provides the necessary background for the development of better diagnostic tools.
回归热疏螺旋体是虱传回归热的病原体,在非洲之角流行。欧洲对此重新引起关注,2015年德国报告的病例数最多(n = 45),其他欧洲国家也有散发病例。到目前为止,仅对一个基因组进行了测序,这阻碍了特异性分子诊断和分型工具的开发。在此,我们报告了回归热疏螺旋体改良的培养条件以及不同年份分离培养的6株菌株的种内基因组变异性,以探索识别新的分型靶点的可能性,并研究该病原体的分子流行病学。
方法/主要发现:来自埃塞俄比亚的2株历史菌株以及2015年从索马里(n = 3)和埃塞俄比亚(n = 1)的移民中获得的4株菌株,在补充有50%胎牛血清的MPK培养基中培养。使用Illumina MiSeq技术对全基因组进行测序,并使用CLC基因组工作台和SPAdes从头组装软件进行分析。与参考回归热疏螺旋体A1相比,在分布于染色体和质粒上的基因组中鉴定出29 - 38个单核苷酸多态性。此外,还鉴定出与现有参考基因组相比长度不同的质粒。
结论/意义:观察到的回归热疏螺旋体菌株低遗传变异性可能是由于其适应了非常保守的媒介 - 宿主(虱子 - 人类)循环,或者受到病原体苛求的特性及其对体外生长的抗性的影响。然而,2015年获得的菌株具有相同的染色体单核苷酸多态性,通过全基因组测序可与历史菌株区分开来,但不能通过迄今使用的分型方法区分。这是第一项研究回归热疏螺旋体分子流行病学的研究,并为开发更好的诊断工具提供了必要的背景。