Walker Melanie, Uranga Carla, Levy Samuel Hs, Kelly Cory, Edlund Anna
Department of Neurological Surgery, University of Washington, Seattle, Washington, United States.
Genomic Medicine Group, J. Craig Venter Institute, La Jolla, California, United States.
Surg Neurol Int. 2022 Jun 10;13:247. doi: 10.25259/SNI_104_2022. eCollection 2022.
Despite a reduction in stroke incidence and age-standardized death rates, stroke remains a leading cause of death and disability worldwide. Significant interest in recent years has focused on the microbiota-host interaction because accumulating evidence has revealed myriad ways in which bacteria may contribute to risk of stroke and adverse outcomes after stroke. The emergence of endovascular thrombectomy as a treatment provides a unique opportunity to utilize thrombus retrieved from cerebral arteries to fill knowledge gaps about the influence of bacteria on stroke pathophysiology. While bacterial signatures have been confirmed in cerebral thrombi, the exact nature of the pathogenesis has not been established.
Thrombi were obtained from a cohort of adult ischemic stroke patients during standard of care thrombectomy. After DNA extraction and quantification, thrombi underwent 16S rRNA amplicon-based metagenomic sequencing, followed by bioinformatics processing. Taxonomic identification of bacterial colonies isolated on Agar plates from plated suspension was performed using DNA extraction and full length 16S Sanger sequencing.
A broad diversity of bacterial signatures was identified in specimens, primarily of cariogenic origin.
In this small study, we demonstrate proof of concept and technical feasibility for amplicon-based metagenomic sequencing of arterial thrombi and briefly discuss preliminary findings, challenges, and near-term translational opportunities for thrombus genomics.
尽管中风发病率和年龄标准化死亡率有所下降，但中风仍是全球主要的死亡和残疾原因。近年来，人们对微生物群与宿主的相互作用产生了浓厚兴趣，因为越来越多的证据表明，细菌可能通过多种方式导致中风风险及中风后的不良后果。血管内血栓切除术作为一种治疗方法的出现，为利用从脑动脉中取出的血栓来填补关于细菌对中风病理生理学影响的知识空白提供了独特机会。虽然已经在脑血栓中证实了细菌特征，但发病机制的确切性质尚未明确。
在标准治疗性血栓切除术期间，从一组成年缺血性中风患者中获取血栓。提取DNA并进行定量后，对血栓进行基于16S rRNA扩增子的宏基因组测序，随后进行生物信息学处理。使用DNA提取和全长16S桑格测序对从平板悬浮液接种在琼脂平板上分离出的细菌菌落进行分类鉴定。
在标本中鉴定出种类繁多的细菌特征，主要源自致龋菌。
在这项小型研究中，我们证明了对动脉血栓进行基于扩增子的宏基因组测序的概念验证和技术可行性，并简要讨论了血栓基因组学的初步发现、挑战和近期转化机会。
