Genomic Analyses of Acute Flaccid Myelitis Cases among a Cluster in Arizona Provide Further Evidence of Enterovirus D68 Role
Genomic Analyses of Acute Flaccid Myelitis Cases among a Cluster in Arizona Provide Further Evidence of Enterovirus D68 Role
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DOI：
10.1128/mbio.02262-18
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发表时间：
2019-01-01
期刊：
影响因子：
6.4
通讯作者：
Engelthaler, David M.
中科院分区：
文献类型：
--
作者：
Bowers, Jolene R.;Valentine, Michael;Engelthaler, David M.
肠道病毒是呼吸道和胃肠道疾病的常见原因，包括脊髓灰质炎病毒在内的多种亚型可引起神经系统疾病。近年来，肠道病毒 D68 (EV-D68) 与严重的神经系统疾病有关，包括急性弛缓性脊髓炎 (AFM)，通常先有呼吸道疾病。 2016 年 9 月，亚利桑那州菲尼克斯发现了 11 起儿童 AFM 疑似病例。为了确定这些病例是否与 EV-D68 相关，我们对患者的鼻咽 (NP) 拭子和脑脊液 (CSF) 材料进行了多项基因组分析，包括针对 EV-D68 VP1 基因的实时 PCR 和扩增子测序以及公正的微生物组和宏基因组测序。 11 名患者中有 4 名被归类为 AFM 确诊病例，另有 1 名病例被归类为 AFM 疑似病例。实时 PCR 和扩增子测序在收集 NP 拭子的三名 AFM 患者以及第四名被诊断患有急性播散性脑脊髓炎的患者中检测到了 EV-D68 病毒 RNA，这种疾病通常发生在细菌或病毒感染（包括肠道病毒）之后。在这些病例中，16S 或 RNA 和 DNA 宏基因组测序未发现 AFM 的其他明显病因，这增强了 EV-D68 是病因的可能性。在第四例 AFM 病例和该集群中的另一例可疑病例的脑脊液中检测到单纯疱疹病毒 DNA。多种基因组技术（例如此处描述的技术）可用于诊断疑似 EV-D68 呼吸道疾病的患者、帮助 AFM 诊断以及未来的 EV-D68 监测和流行病学。 重要性 肠道病毒经常导致呼吸道和胃肠道疾病；然而，包括脊髓灰质炎病毒在内的多种亚型可导致严重的神经系统疾病。最近两年（即 2014 年、2016 年和 2018 年）非脊髓灰质炎急性弛缓性麻痹病例的增加引发了人们的猜测：其他肠道病毒，特别是肠道病毒 D68 (EV-D68) 正在出现，以填补根除脊髓灰质炎病毒留下的空白。 2016 年，亚利桑那州菲尼克斯发现了 11 起疑似儿童急性弛缓性脊髓炎 (AFM) 病例。多重基因组分析发现大多数临床 AFM 病例中都存在 EV-D68。除了有限地检测到疱疹病毒外，该集群中没有发现其他可能的病因。这些发现增强了 EV-D68 是 AFM 病因的可能性，并表明为本研究开发的快速分子检测对于 AFM 和 EV-D68 的研究很有用。
Enteroviruses are a common cause of respiratory and gastrointestinal illness, and multiple subtypes, including poliovirus, can cause neurologic disease. In recent years, enterovirus D68 (EV-D68) has been associated with serious neurologic illnesses, including acute flaccid myelitis (AFM), frequently preceded by respiratory disease. A cluster of 11 suspect cases of pediatric AFM was identified in September 2016 in Phoenix, AZ. To determine if these cases were associated with EV-D68, we performed multiple genomic analyses of nasopharyngeal (NP) swabs and cerebrospinal fluid (CSF) material from the patients, including real-time PCR and amplicon sequencing targeting the EV-D68 VP1 gene and unbiased microbiome and metagenomic sequencing. Four of the 11 patients were classified as confirmed cases of AFM, and an additional case was classified as probable AFM. Real-time PCR and amplicon sequencing detected EV-D68 virus RNA in the three AFM patients from which NP swabs were collected, as well as in a fourth patient diagnosed with acute disseminated encephalomyelitis, a disease that commonly follows bacterial or viral infections, including enterovirus. No other obvious etiological causes for AFM were identified by 16S or RNA and DNA metagenomic sequencing in these cases, strengthening the likelihood that EV-D68 is an etiological factor. Herpes simplex viral DNA was detected in the CSF of the fourth case of AFM and in one additional suspect case from the cluster. Multiple genomic techniques, such as those described here, can be used to diagnose patients with suspected EV-D68 respiratory illness, to aid in AFM diagnosis, and for future EV-D68 surveillance and epidemiology.IMPORTANCE Enteroviruses frequently result in respiratory and gastrointestinal illness; however, multiple subtypes, including poliovirus, can cause severe neurologic disease. Recent biennial increases (i.e., 2014, 2016, and 2018) in cases of non-polio acute flaccid paralysis have led to speculations that other enteroviruses, specifically enterovirus D68 (EV-D68), are emerging to fill the niche that was left from poliovirus eradication. A cluster of 11 suspect cases of pediatric acute flaccid myelitis (AFM) was identified in 2016 in Phoenix, AZ. Multiple genomic analyses identified the presence of EV-D68 in the majority of clinical AFM cases. Beyond limited detection of herpesvirus, no other likely etiologies were found in the cluster. These findings strengthen the likelihood that EV-D68 is a cause of AFM and show that the rapid molecular assays developed for this study are useful for investigations of AFM and EV-D68.