#Surveillance of #enteroviruses from #paediatric patients attended at a tertiary #hospital in #Catalonia from 2014 to 2017 (J Clin Virol., abstract)

[Source: Science Direct, full page: (LINK). Abstract, edited.]

Journal of Clinical Virology / Available online 30 November 2018 / In Press, Accepted Manuscript

Surveillance of enteroviruses from paediatric patients attended at a tertiary hospital in Catalonia from 2014 to 2017

Cristina Andrés a, Jorgina Vila b, Laura Gimferrer a , Maria Piñana a, Juliana Esperalba a, Maria Gema Codina a, Meritxell Barnés b, Mariadel Carmen Martín a, Francisco Fuentes a, Susana Rubio a, Pilar Alcubilla a, Carlos Rodrigo b, Tomàs Pumarola a, Andrés Antón a

{a} Respiratory Viruses Unit, Virology Section, Microbiology Department, Hospital Universitari Vall d’Hebron, Vall d’Hebron Research Institute, Universitat Autònoma de Barcelona, Barcelona, Spain; {b} Paediatric Hospitalisation Unit, Department of Paediatrics, Hospital Universitari Maternoinfantil Vall d’Hebron, Universitat Autònoma de Barcelona, Barcelona, Spain

Received 4 September 2018, Revised 26 October 2018, Accepted 16 November 2018, Available online 30 November 2018.

DOI: https://doi.org/10.1016/j.jcv.2018.11.004



  • The study reports virological and clinical enterovirus surveillance in Catalonia.
  • The four enterovirus species cocirculated, distinguishing up to 27 different types.
  • Most of neurological studied cases were from the 2016 spring outbreak.
  • EV-A71 was one of the most detected EV, mostly during the outbreak.
  • Rhombencephalitis cases were related to EV-A71 infection.
  • EV-D68 was associated with lower respiratory tract infections.
  • Necessity to perform EV surveillance in primary care settings.




Enterovirus (EV) infections are usually asymptomatic or mild, but symptomatic infections can evolve to severe complications. Outbreaks of EV-A71 and EV-D68 have been recently reported worldwide, sometimes related to severe clinical outcomes.


To describe EV genetic diversity and the clinical outcomes from paediatric patients attended at a tertiary university hospital in Barcelona (Catalonia, Spain) from 2014 to 2017.

Study design

Specimens were collected from paediatric (<17 years old) cases with suspicion of respiratory tract infection or EV infection. EV laboratory-confirmation was performed by specific real-time multiplex RT-PCR assay. Partial viral VP1 protein was sequenced for genetic characterisation by phylogenetic analyses.


A total of 376 (7%) from 5,703 cases were EV laboratory-confirmed. Phylogenetic analyses of VP1 (210; 81%) sequences distinguished up to 27 different EV types distributed within EV-A (82; 40%), EV-B (90; 42%), EV-C (5; 2%), and EV-D (33; 15%), in addition to 50 (19%) rhinoviruses. The most predominant were EV-A71 (37; 45%) and EV-D68 (32; 99%). EV-A71 was highly related to neurological complications (25/39, 63%), of which 20/39 were rhombencephalitis, and most EV-D68 (28/32, 88%) were associated with lower respiratory tract infections (LRTI), and exceptionally one (3%) with flaccid paralysis.


EV-A71 and EV-D68 were the most detected EV in respiratory specimens. EV-A71 was highly related to neurological disease and EV-D68 was often associated with LRTI. However, both potential relatedness to neurological diseases makes the monitoring of EV circulation obligatory.

Keywords: enteroviruses – respiratory infections – surveillance – genetic diversity – molecular epidemiology – paediatric population

© 2018 Elsevier B.V. All rights reserved.

Keywords: Enterovirus; EV-A71; EV-D68; Rhomboencephalitis; AFP; Spain.



Contemporary Circulating #Enterovirus D68 Strains Have Acquired the Capacity for Viral Entry and #Replication in #Human #Neuronal Cells (mBio, abstract)

[Source: mBio, full page: (LINK). Abstract, edited.]

Contemporary Circulating Enterovirus D68 Strains Have Acquired the Capacity for Viral Entry and Replication in Human Neuronal Cells

David M. Brown, Alison M. Hixon, Lauren M. Oldfield, Yun Zhang, Mark Novotny, Wei Wang, Suman R. Das, Reed S. Shabman, Kenneth L. Tyler, Richard H. Scheuermann

Diane E. Griffin, Editor

DOI: 10.1128/mBio.01954-18



Enterovirus D68 (EV-D68) has historically been associated with respiratory illnesses. However, in the summers of 2014 and 2016, EV-D68 outbreaks coincided with a spike in polio-like acute flaccid myelitis/paralysis (AFM/AFP) cases. This raised concerns that EV-D68 could be the causative agent of AFM during these recent outbreaks. To assess the potential neurotropism of EV-D68, we utilized the neuroblastoma-derived neuronal cell line SH-SY5Y as a cell culture model to determine if differential infection is observed for different EV-D68 strains. In contrast to HeLa and A549 cells, which support viral infection of all EV-D68 strains tested, SH-SY5Y cells only supported infection by a subset of contemporary EV-D68 strains, including isolates from the 2014 outbreak. Viral replication and infectivity in SH-SY5Y were assessed using multiple assays: virus production, cytopathic effects, cellular ATP release, and VP1 capsid protein production. Similar differential neurotropism was also observed in differentiated SH-SY5Y cells, primary human neuron cultures, and a mouse paralysis model. Using the SH-SY5Y cell culture model, we determined that barriers to viral binding and entry were at least partly responsible for the differential infectivity phenotype. Transfection of genomic RNA into SH-SY5Y generated virions for all EV-D68 isolates, but only a single round of replication was observed from strains that could not directly infect SH-SY5Y. In addition to supporting virus replication and other functional studies, this cell culture model may help identify the signatures of virulence to confirm epidemiological associations between EV-D68 strains and AFM and allow for the rapid identification and characterization of emerging neurotropic strains.



Since the EV-D68 outbreak during the summer of 2014, evidence of a causal link to a type of limb paralysis (AFM) has been mounting. In this article, we describe a neuronal cell culture model (SH-SY5Y cells) in which a subset of contemporary 2014 outbreak strains of EV-D68 show infectivity in neuronal cells, or neurotropism. We confirmed the difference in neurotropism in vitro using primary human neuron cell cultures and in vivo with a mouse paralysis model. Using the SH-SY5Y cell model, we determined that a barrier to viral entry is at least partly responsible for neurotropism. SH-SY5Y cells may be useful in determining if specific EV-D68 genetic determinants are associated with neuropathogenesis, and replication in this cell line could be used as rapid screening tool for identification of neurotropic EV-D68 strains. This may assist with better understanding of pathogenesis and epidemiology and with the development of potential therapies.

Keywords: EV-D78; AFM; AFP.


#Investigation of #AFP Reported with #LaCrosse Virus #Infection, #Ohio, #USA, 2008–2014 (Emerg Infect Dis., abstract)

[Source: US Centers for Disease Control and Prevention (CDC), Emerging Infectious Diseases Journal, full page: (LINK). Abstract, edited.]

Volume 23, Number 12—December 2017 / Dispatch

Investigation of Acute Flaccid Paralysis Reported with La Crosse Virus Infection, Ohio, USA, 2008–2014

Morgan J. Hennessey, Daniel M. Pastula, Kimberly Machesky, Marc Fischer, Nicole P. Lindsey, Mary DiOrio, J. Erin Staples  , and Sietske de Fijter

Author affiliations: Centers for Disease Control and Prevention, Atlanta, Georgia, USA (M.J. Hennessey, D.M. Pastula); Centers for Disease Control and Prevention, Fort Collins, Colorado, USA (M.J. Hennessey, D.M. Pastula, M. Fischer, N.P. Lindsey, J.E. Staples); Ohio Department of Health, Columbus, Ohio, USA (K. Machesky, M. DiOrio, S. de Fijter)



Infection with La Crosse virus can cause meningoencephalitis, but it is not known to cause acute flaccid paralysis (AFP). During 2008–2014, nine confirmed or probable La Crosse virus disease cases with possible AFP were reported in Ohio, USA. After an epidemiologic and clinical investigation, we determined no patients truly had AFP.

Keywords: La Crosse Virus; AFP; USA.


Co-circulation of multiple subtypes of #enterovirus A71 (EV- A71) genotype C, including novel #recombinants characterised by use of whole genome sequencing (WGS), #Denmark 2016 (@eurosurveillanc, abstract)

[Source: Eurosurveillance, full page: (LINK). Abstract, edited.]

Eurosurveillance, Volume 22, Issue 26, 29 June 2017  / Rapid communication

Co-circulation of multiple subtypes of enterovirus A71 (EV- A71) genotype C, including novel recombinants characterised by use of whole genome sequencing (WGS), Denmark 2016

SE Midgley 1 , AG Nielsen 1 , R Trebbien 1 , MW Poulsen 1 , PH Andersen 2 , TK Fischer 1 3

Author affiliations: 1. Section for Virus Surveillance and Research, Statens Serum Institut, Copenhagen, Denmark; 2. Infectious Disease Epidemiology, Statens Serum Institut, Copenhagen, Denmark; 3. Center for Global Health, Department of Infectious Diseases, University of Southern Denmark, Odense, Denmark

Correspondence: Sofie Elisabeth Midgley (soi@ssi.dk)

Citation style for this article: Midgley SE, Nielsen AG, Trebbien R, Poulsen MW, Andersen PH, Fischer TK. Co-circulation of multiple subtypes of enterovirus A71 (EV- A71) genotype C, including novel recombinants characterised by use of whole genome sequencing (WGS), Denmark 2016. Euro Surveill. 2017;22(26):pii=30565. DOI: http://dx.doi.org/10.2807/1560-7917.ES.2017.22.26.30565

Received:19 May 2017; Accepted:29 June 2017



In Europe, enterovirus A71 (EV-A71) has primarily been associated with sporadic cases of neurological disease. The recent emergence of new genotypes and larger outbreaks with severely ill patients demonstrates a potential for the spread of new, highly pathogenic EV-A71 strains. Detection and characterisation of these new emerging EV variants is challenging as standard EV assays may not be adequate, necessitating the use of whole genome analysis.

Keywords: EV-A71; Enterovirus; Denmark; HFMD; Acute Flaccid Paralysis.


#Crystal #structure and #thermostability characterization of #EVD68 3Dpol (J Virol., abstract)

[Source: Journal of Virology, full page: (LINK). Abstract, edited.]

Crystal structure and thermostability characterization of EV-D68-3Dpol

Chunnian Wang1,3, Caiyan Wang2,3, Qing Li1,3, Zhong Wang1,3* and Wei Xie2,3*

Author Affiliations: 1School of Pharmaceutical Sciences, The Sun Yat-Sen University, 132 E. Circle Rd. University City, Guangzhou, Guangdong 510006, People’s Republic of China. 2State Key Laboratory for Biocontrol, School of Life Sciences, The Sun Yat-Sen University, 135 W. Xingang Rd., Guangzhou, Guangdong 510275, People’s Republic of China. 3Center for Cellular & Structural biology, The Sun Yat-Sen University, 132 E. Circle Rd., University City, Guangzhou, Guangdong 510006, People’s Republic of China.



Enterovirus-D68 (EV-D68) is one of the many non-polio enteroviruses that cause mild-to-severe respiratory illness. The non-structural protein 3Dpol is an RNA-dependent RNA polymerase (RdRP) of EV-D68, which plays a critical role in the replication of the viral genome and represents a promising drug target. Here we report the first three-dimensional crystal structure of RdRP from EV-D68 in complex with the substrate GTP to 2.3-Å resolution. The RdRP structure is similar to those of other viral RdRPs, where the three domains termed the palm, fingers, and thumb form a structure resembling a cupped right hand. Particularly, an N-terminal fragment (Gly1-Phe30) bridges the fingers and the thumb domains, which accounts for the enhanced stability of the full-length enzyme over the truncation mutant, as assessed by our thermal shift assays and the dynamic light scattering studies. Additionally, the GTP molecule bound proximal to the active site interacts with both the palm and fingers domains to stabilize the core structure of 3Dpol. Interestingly, using limited proteolysis assays, we found that different NTPs stabilize the polymerase structure by various degrees, with GTP and CTP being the most and least stabilizing nucleosides respectively. Lastly, we derived a model of the core structure of 3Dpol stabilized by GTP, according to our proteolytic studies. The biochemical and biophysical characterization conducted in this study helps us to understand the stability of EV-D68-3Dpol, which may extend to other RdRPs as well.



Enterovirus D68 (EV-D68) is an emerging viral pathogen, which caused sporadic infections around the world. In recent years, epidemiology studies have reported an increasing number of patients with respiratory diseases globally due to the EV-D68 infection. Moreover, the infection has been associated with acute flaccid paralysis and cranial nerve dysfunction in children. However, there are no vaccines and antiviral treatments specifically targeting the virus to date. In this study, we solved the crystal structure of the RNA-dependent RNA polymerase of EV-D68, and carried out systematic biophysical and biochemical characterization on the overall and local structural stability of the WT enzyme and several variants, which yields a clear view on the structure-activity relaitionship of the EV-D68 RNA polymerase.



*: Correspondence: Wei Xie: xiewei6@mail.sysu.edu.cn and Zhong Wang: wangzh357@mail.sysu.edu.cn.

Copyright © 2017 American Society for Microbiology. All Rights Reserved.

Keywords: Enterovirus; EV-D68; HFMD; Acute Flaccid Paralysis.


#Enteroviral T-cell #encephalitis related to immunosuppressive therapy including #rituximab (Neurology, abstract)

[Source: Neurology, full page: (LINK). Abstract, edited.]

Enteroviral T-cell encephalitis related to immunosuppressive therapy including rituximab

Nicolas Garzo-Caldas, MD, Elena Ruiz-Sainz, MD, Sara Vila-Bedmar, MD, Sara Llamas-Velasco, MD, Aurelio Hernández-Lain, MD, PhD, Juan Ruiz-Morales, MD, Dolores Folgueira-López, MD, PhD and Alberto Villarejo-Galende, MD, PhD

AFFILIATIONS: From Hospital Universitario 12 de Octubre (N.G.-C., S.V.-B., S.L.-V., A.H.-L., J.R.-M., D.F.-L., A.V.-G.); CIBERNED (S.L.-V., A.V.-G.), Madrid; and Hospital del Tajo (E.R.-S.), Aranjuez, Spain.

Correspondence to Dr. Villarejo-Galende: avgalende@yahoo.es

Published online before print June 23, 2017, doi: http:/​/​dx.​doi.​org/​10.​1212/​WNL.​0000000000004148

Neurology 10.1212/WNL.0000000000004148



Enteroviruses are a common cause of aseptic meningitis and mild infections in childhood. However, in immunocompromised patients they may cause severe neurologic conditions such as encephalitis or polio-like paralysis.

Received October 27, 2016. Accepted in final form April 19, 2017.

© 2017 American Academy of Neurology

Keywords: Enterovirus; Acute Flaccid Paralysis; Encephalitis.


Acute segmental #poliomyelitis-like flaccid #paralysis in an #adult in the #UK, associated with #EVD68 (Pract Neurol., abstract)

[Source: US National Library of Medicine, full page: (LINK). Abstract, edited.]

Pract Neurol. 2017 Jun 16. pii: practneurol-2017-001609. doi: 10.1136/practneurol-2017-001609. [Epub ahead of print]

Acute segmental poliomyelitis-like flaccid paralysis in an adult in the UK, associated with enterovirus D68.

Stacpoole SRL1,2, Molyneux A2, Bäumer D1,2.

Author information: 1 Department of Neurology, Peterborough City Hospital, Peterborough and Stamford Hospitals NHS Foundation Trust, Peterborough, UK. 2 Department of Neurology, Addenbrooke’s Hospital, Cambridge University Hospitals NHS Foundation Trust, Cambridge, UK.



Enterovirus D68 has been associated with a poliomyelitis-like illness, notably during an outbreak in 2014, and particularly affecting children in the USA. We report a case of acute segmental flaccid paralysis with respiratory involvement in an adult in the UK, with enterovirus D68 detected in a sputum sample. MR imaging of cervical spinal cord showed a longitudinally extensive T2 hyperintensity in the anterior cord. Cerebrospinal fluid showed an elevated white cell count, predominantly lymphocytic, with otherwise normal constituents and negative viral PCRs. His respiratory function improved after intravenous immunoglobulin, suggesting that this may be useful in such cases. Clinicians should consider enterovirus D68 infection in the differential diagnosis of Guillain-Barré syndrome, particularly the pharyngeal-cervical-brachial variant.

© Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2017. All rights reserved. No commercial use is permitted unless otherwise expressly granted.


PMID: 28626021 DOI: 10.1136/practneurol-2017-001609

Keywords: EV-D68; Enterovirs; Acute Flaccid Paralysis; Poliomyelitis.