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Published on: December 2, 2012
Genomic and serologic characterization of enterovirus A71 brainstem encephalitis
Kristoffer E Leon1, Ryan D Schubert1, Didac Casas-Alba1
1From 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.
Metagenomic next-generation sequencing (mNGS) and CSF pan-viral serology (VirScan) improved detection of enterovirus A71 (EV-A71) in pediatric encephalitis cases. These advanced methods identified more viral infections than conventional tests, aiding in outbreak investigation.
Area of Science:
- Neurovirology
- Infectious Disease Epidemiology
- Genomic Surveillance
Background:
- A 2016 pediatric brainstem encephalitis outbreak in Catalonia was linked to enterovirus A71 (EV-A71).
- Conventional diagnostic methods showed limited sensitivity for detecting EV in cerebrospinal fluid (CSF).
Purpose of the Study:
- To enhance viral detection and characterization during the EV-A71 pediatric encephalitis outbreak.
- To compare the efficacy of metagenomic next-generation sequencing (mNGS) and CSF pan-viral serology (VirScan) against conventional methods.
Main Methods:
- RNA extraction and mNGS analysis of CSF, plasma, stool, and nasopharyngeal samples from affected children.
- CSF samples were analyzed using VirScan, with validation by ELISA.
- Phylogenetic analysis was performed on detected EV-A71 strains.
Main Results:
- mNGS increased EV detection rates compared to qRT-PCR, particularly in previously negative samples.
- VirScan combined with ELISA significantly improved viral detection in CSF compared to mNGS and qRT-PCR alone.
- Phylogenetic analysis linked the outbreak strain to a neurovirulent German EV-A71, with a specific VP1 protein substitution (S241P) found exclusively in the CNS.
Conclusions:
- mNGS and VirScan offer superior sensitivity for detecting EVs in the central nervous system (CNS) compared to qRT-PCR.
- These advanced techniques provide valuable insights into viral immune responses and genomic characteristics during outbreaks.
- The identified S241P substitution may be relevant for EV-A71 neuroinvasion.
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