UNASSIGNED: The overarching objectives of this study were to: 1) describe the distribution of enteroviruses in Arizona during the late summer and fall of 2022, the time of year when they are thought to be most abundant, and 2) demonstrate the utility of viral pan-assay approaches for semi-agnostic discovery that can be followed up by more targeted assays and phylogenomics.
UNASSIGNED: This study utilizes pooled nasal samples collected from school-aged children and long-term care facility residents, and wastewater from multiple locations in Arizona during July-October of 2022. We used PCR to amplify and sequence a region common to all enteroviruses, followed by species-level bioinformatic characterization using the QIIME 2 platform. For Enterovirus-D68 (EV-D68), detection was carried out using RT-qPCR, followed by confirmation using near-complete whole EV-D68 genome sequencing using a newly designed tiled amplicon approach.
UNASSIGNED: In the late summer and early fall of 2022, multiple enterovirus species were identified in Arizona wastewater, with Coxsackievirus A6, EV-D68, and Coxsackievirus A19 composing 86% of the characterized reads sequenced. While EV-D68 was not identified in pooled human nasal samples, and the only reported acute flaccid myelitis case in Arizona did not test positive for the virus, an in-depth analysis of EV-D68 in wastewater revealed that the virus was circulating from August through mid-October. A phylogenetic analysis on this relatively limited dataset revealed just a few importations into the state, with a single clade indicating local circulation.
UNASSIGNED: This study further supports the utility of wastewater-based epidemiology to identify potential public health threats. Our further investigations into EV-D68 shows how these data might help inform healthcare diagnoses for children presenting with concerning neurological symptoms.
■这项研究的总体目标是:1)描述2022年夏末和秋季亚利桑那州肠道病毒的分布,这一年中它们被认为是最丰富的。和2)证明了病毒泛分析方法用于半不可知论发现的实用性,可以通过更有针对性的测定和系统基因组学进行跟进。
■本研究利用从学龄儿童和长期护理机构居民收集的合并鼻样本,以及2022年7月至10月亚利桑那州多个地点的废水。我们使用PCR对所有肠道病毒共有的区域进行扩增和测序,然后使用QIIME2平台进行物种水平的生物信息学表征。对于肠道病毒D68(EV-D68),使用RT-qPCR进行检测,然后使用新设计的平铺扩增子方法使用接近完全的全EV-D68基因组测序进行确认。
■在2022年夏末和初秋,在亚利桑那州的废水中发现了多种肠道病毒物种,柯萨奇病毒A6,EV-D68和柯萨奇病毒A19构成了86%的测序特征读数。虽然在合并的人鼻样本中未发现EV-D68,亚利桑那州唯一报告的急性弛缓性脊髓炎病例没有病毒检测呈阳性,对废水中EV-D68的深入分析显示,该病毒从8月到10月中旬一直在传播。对这个相对有限的数据集进行的系统发育分析显示,该州只有一些输入,有一个分支指示局部循环。
■这项研究进一步支持基于废水的流行病学的实用性,以确定潜在的公共卫生威胁。我们对EV-D68的进一步调查显示,这些数据可能有助于为出现神经系统症状的儿童提供医疗诊断。