Department of Biological Sciences, Hunter College of the City University of New York, New York, New York, USA.
Graduate Center of the City University of New York, New York, New York, USA.
J Clin Microbiol. 2018 Oct 25;56(11). doi: 10.1128/JCM.00940-18. Print 2018 Nov.
A mixed infection of a single tick or host by Lyme disease spirochetes is common and a unique challenge for the diagnosis, treatment, and surveillance of Lyme disease. Here, we describe a novel protocol for differentiating Lyme strains on the basis of deep sequencing of the hypervariable outer surface protein C locus (). Improving upon the traditional DNA-DNA hybridization method, the next-generation sequencing-based protocol is high throughput, quantitative, and able to detect new pathogen strains. We applied the method to more than one hundred infected ticks collected from New York State, USA, in 2015 and 2016. An analysis of strain distributions within individual ticks suggests an overabundance of multiple infections by five or more strains, inhibitory interactions among coinfecting strains, and the presence of a new strain closely related to A supporting bioinformatics pipeline has been developed. The newly designed pair of universal primers target intergenic sequences conserved among all known Lyme pathogens. The protocol could be used for culture-free identification and quantification of Lyme pathogens in wildlife and potentially in clinical specimens.
莱姆病螺旋体的单一蜱虫或宿主的混合感染很常见，这对莱姆病的诊断、治疗和监测是一个独特的挑战。在这里，我们描述了一种基于超变外壳蛋白 C 基因座（）的深度测序来区分莱姆菌株的新方案。与传统的 DNA-DNA 杂交方法相比，基于下一代测序的方案具有高通量、定量和能够检测新病原体菌株的能力。我们将该方法应用于 2015 年和 2016 年从美国纽约州收集的一百多个感染的蜱虫。对单个蜱虫内菌株分布的分析表明，存在多种菌株的过度感染（超过五种）、共感染菌株之间的抑制相互作用，以及一种与 密切相关的新菌株的存在。已开发出支持的生物信息学管道。新设计的一对通用引物针对所有已知莱姆病原体中保守的基因间序列。该方案可用于野生动物中莱姆病原体的无培养鉴定和定量，并且有可能用于临床标本。
