肠道病毒 A71 脑干脑炎的基因组和血清学特征。
Genomic and serologic characterization of enterovirus A71 brainstem encephalitis.
机构信息
From the Medical Scientist Training Program (K.E.L.), University of California, San Francisco; Biomedical Sciences Graduate Program (K.E.L., I.A.H.), University of California, San Francisco; Weill Institute for Neurosciences (R.D.S., I.A.H., P.S.R., A.R., M.R.W.), University of California, San Francisco; Department of Neurology (R.D.S., I.A.H., P.S.R., A.R., M.R.W.), University of California, San Francisco; Institut de Recerca Pediàtrica Hospital Sant Joan de Déu (D.C.-A., C.L., A.V.-R., C.M.-A.), Barcelona, Spain; Chan Zuckerberg Biohub (J.E.P., W.W., C.K.C., E.D.C., J.L.D.), San Francisco; Department of Biochemistry and Biophysics (L.M.K., H.A.S., K.C.Z., J.L.D.), University of California, San Francisco; CIBER Epidemiología y Salud Pública (CIBERESP) (C.L., M.C., C.M.-A.), Health Institute Carlos III; Department of Pediatrics (C.L.), Universitat de Barcelona, Barcelona; Enterovirus Unit (M.C.), Spanish National Centre for Microbiology, Instituto de Salud Carlos III, Madrid, Spain; Division of Infectious Diseases (C.L.), Department of Medicine, University of California, San Francisco; and Department of Medicine. Universitat Internacional de Catalunya (C.M.-A.), Barcelona, Spain.
出版信息
Neurol Neuroimmunol Neuroinflamm. 2020 Mar 5;7(3). doi: 10.1212/NXI.0000000000000703. Print 2020 May.
OBJECTIVE
In 2016, Catalonia experienced a pediatric brainstem encephalitis outbreak caused by enterovirus A71 (EV-A71). Conventional testing identified EV in the periphery but rarely in CSF. Metagenomic next-generation sequencing (mNGS) and CSF pan-viral serology (VirScan) were deployed to enhance viral detection and characterization.
METHODS
RNA was extracted from the CSF (n = 20), plasma (n = 9), stool (n = 15), and nasopharyngeal samples (n = 16) from 10 children with brainstem encephalitis and 10 children with meningitis or encephalitis. Pathogens were identified using mNGS. Available CSF from cases (n = 12) and pediatric other neurologic disease controls (n = 54) were analyzed with VirScan with a subset (n = 9 and n = 50) validated by ELISA.
RESULTS
mNGS detected EV in all samples positive by quantitative reverse transcription polymerase chain reaction (qRT-PCR) (n = 25). In qRT-PCR-negative samples (n = 35), mNGS found virus in 23% (n = 8, 3 CSF samples). Overall, mNGS enhanced EV detection from 42% (25/60) to 57% (33/60) (-value = 0.013). VirScan and ELISA increased detection to 92% (11/12) compared with 46% (4/12) for CSF mNGS and qRT-PCR (-value = 0.023). Phylogenetic analysis confirmed the EV-A71 strain clustered with a neurovirulent German EV-A71. A single amino acid substitution (S241P) in the EVA71 VP1 protein was exclusive to the CNS in one subject.
CONCLUSION
mNGS with VirScan significantly increased the CNS detection of EVs relative to qRT-PCR, and the latter generated an antigenic profile of the acute EV-A71 immune response. Genomic analysis confirmed the close relation of the outbreak EV-A71 and neuroinvasive German EV-A71. A S241P substitution in VP1 was found exclusively in the CSF.
目的
2016 年，加泰罗尼亚遭遇由肠道病毒 A71（EV-A71）引起的小儿脑干脑炎疫情。常规检测在外周血中发现了 EV，但在脑脊液中很少发现。采用宏基因组下一代测序（mNGS）和脑脊液泛病毒血清学（VirScan）来提高病毒检测和鉴定能力。
方法
从 10 例脑干脑炎患儿和 10 例脑膜炎或脑炎患儿的脑脊液（n=20）、血浆（n=9）、粪便（n=15）和鼻咽样本（n=16）中提取 RNA。采用 mNGS 鉴定病原体。对病例（n=12）和儿科其他神经疾病对照（n=54）的可用脑脊液进行 VirScan 分析，并用 ELISA 对其中部分样本（n=9 和 n=50）进行验证。
结果
mNGS 在所有经定量逆转录聚合酶链反应（qRT-PCR）检测呈阳性的样本（n=25）中均检测到 EV。在 qRT-PCR 阴性样本（n=35）中，mNGS 检出病毒的比例为 23%（n=8，3 例脑脊液样本）。总体而言，mNGS 将 EV 的检出率从 42%（25/60）提高到 57%（33/60）（-值=0.013）。与脑脊液 mNGS 和 qRT-PCR 的 46%（4/12）相比，VirScan 和 ELISA 将 EV 的检出率提高到 92%（11/12）（-值=0.023）。系统进化分析证实，分离株与德国神经侵袭性 EV-A71 密切相关。在 1 名患者的 CNS 中发现 EV-A71 VP1 蛋白中的单个氨基酸取代（S241P）是独特的。
结论
与 qRT-PCR 相比，mNGS 联合 VirScan 可显著提高 CNS 中 EV 的检出率，后者可产生急性 EV-A71 免疫反应的抗原谱。基因组分析证实了暴发的 EV-A71 与神经侵袭性德国 EV-A71 密切相关。VP1 中的 S241P 取代仅在脑脊液中发现。
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