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. 2020 Mar 2;10:3810. doi: 10.1038/s41598-020-60761-5

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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EV-A71 viral isolates from 2006 outbreak in Sarawak, Malaysia, and its viral kinetics in human rhabdomyosarcoma (RD) and neuroblastoma (SH-SY5Y) cells. (a) Clinical information about the viruses isolated including genotype, symptoms and area virus was isolated from. (b–e) RD and SH-SY5Y cells were infected with mild, moderate and severe isolates of EV-A71 at MOI 10 and 1 respectively, and harvested at 0, 3, 6, 12 and 24 hours post-infection (hpi). (b,c) Infection kinetics in RD cells via (b) quantification of VP1 antigen by flow cytometry and (c) viral load quantification of VP1 RNA by qRT-PCR. (d,e) Infection kinetics in SH-SY5Y cells via (d) quantification of VP1 antigen by flow cytometry and (e) viral load quantification of VP1 RNA by qRT-PCR. Data are presented as mean ± SEM and representative of three independent experiments. Statistical analysis was carried out with Kruskal-Wallis with Dunn’s multiple comparisons test to compare among EV-A71 isolates at the respective time-points (**p<0.01; ***p<0.001) [Mild vs Severe EV-A71 (*); Moderate vs Severe EV-A71 (+)].