Title Detection and Typing of Human Enteroviruses from Clinical Samples by Entire-Capsid Next Generation Sequencing.
Author Majumdar, Manasi; Celma, Cristina; Pegg, Elaine; Polra, Krunal; Dunning, Jake; Martin, Javier
Journal Viruses Publication Year/Month 2021-Apr
PMID 33918088 PMCID PMC8070635
Affiliation + expend 1.Division of Virology, National Institute for Biological Standards and Control (NIBSC), Potters Bar, Hertfordshire EN6 3QG, UK.

There are increasing concerns of infections by enteroviruses (EVs) causing severe disease in humans. EV diagnostic laboratory methods show differences in sensitivity and specificity as well as the level of genetic information provided. We examined a detection method for EVs based on next generation sequencing (NGS) analysis of amplicons covering the entire capsid coding region directly synthesized from clinical samples. One hundred and twelve clinical samples from England; previously shown to be positive for EVs, were analyzed. There was high concordance between the results obtained by the new NGS approach and those from the conventional Sanger method used originally with agreement in the serotypes identified in the 83 samples that were typed by both methods. The sensitivity and specificity of the NGS method compared to those of the conventional Sanger sequencing typing assay were 94.74% (95% confidence interval, 73.97% to 99.87%) and 97.85% (92.45% to 99.74%) for Enterovirus A, 93.75% (82.80% to 98.69%) and 89.06% (78.75% to 95.49%) for Enterovirus B, 100% (59.04% to 100%) and 98.10% (93.29% to 99.77%) for Enterovirus C, and 100% (75.29% to 100%) and 100% (96.34% to 100%) for Enterovirus D. The NGS method identified five EVs in previously untyped samples as well as additional viruses in some samples, indicating co-infection. This method can be easily expanded to generate whole-genome EV sequences as we show here for EV-D68. Information from capsid and whole-genome sequences is critical to help identifying the genetic basis for changes in viral properties and establishing accurate spatial-temporal associations between EV strains of public health relevance.

StrainID RV Species Serotype Length(nt) Country Year Strain Name
CNP01222 C 42 3886 United Kingdom 2018 CLI-B3-21-RhinoC
CNS01223 C None 2491 United Kingdom 2018 CLI-B3-14-RhinoC
CNZ01221 C 15 3949 United Kingdom 2018 CLI-B3-70-RhinoC
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