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00925njm 22002777a 4500 dc Couderé, Karen author Benschop, Kimberley SM author Koen, Gerrit author van Eijk, Hetty author Harvala, Heli author Bailly, Jean-Luc author Bessaud, Maël author Kamau, Everlyn author Simon, Isabelle author Joffret, Marie-Line author Nikolaeva-Glomb, Lubomira author Georgieva, Irina author Stoyanova, Asya author Diedrich, Sabine author Böttcher, Sindy author Cabrerizo, Maria author Tabain, Irena author Hruškar, Željka author Stevanović, Vladimir author Susi, Petri author Hietanen, Eero author Palminha, Paula author Rainetová, Petra author Baicus, Anda author Kristensen, Maartje author PajkrtJean-Luc Murk, Dasja author Wolthers, Katja C author European Non-Polio Enterovirus Network author 2024-09 Non-polio enteroviruses (NPEV) cause significant disease worldwide. Population-based sero-surveillance, by measuring antibodies against specific NPEV types, provides additional information on past circulation and the prediction for future upsurges. Virus neutralisation assays (VNA), the current method of choice for measuring NPEV type specific antibodies, are not entirely standardised. Via the European Non-Polio Enterovirus Network, we organised a VNA quality assessment in which twelve laboratories participated. We provided five echovirus (E) types (E1, E18, E30 G2, E30 G6 and E6) and intravenous immunoglobulins (IVIG) as a sample for the NPEV VNA quality assessment. Differences in VNA protocols and neutralising Ab (nAb) titres were found between the participating laboratories with geometric coefficients of variation ranging from 10.3-62.9 %. Mixed-effects regression analysis indicated a small but significant effect of type of cell line used. Harmonisation of cell line passage number, however, did not improve variation between laboratories. Calibration by making use of a reference sample, reduced variation between laboratories but differences in nAb titres remained higher than two log2 dilution steps. In conclusion, sero-surveillance data from different laboratories should be compared with caution and standardised protocols are needed. Couderé K, Benschop KSM, Koen G, van Eijk H, Harvala H, Bailly JL, Bessaud M, Kamau E, Simon I, Joffret ML, Nikolaeva-Glomb L, Georgieva I, Stoyanova A, Diedrich S, Böttcher S, Cabrerizo M, Tabain I, Hruškar Ž, Stevanović V, Susi P, Hietanen E, Palminha P, Rainetová P, Baicus A, Kristensen M, Pajkrt D, Murk JL, Wolthers KC, European Non-Polio Enterovirus Network. Assessment of twelve echovirus virus-neutralisation assays in Europe: recommendations for harmonisation of non-polio enterovirus sero-surveillance studies. J Gen Virol. 2024 Sep;105(9). 10.1099/jgv.0.001986 1465-2099 0022-1317 The Journal of general virology 39311842 https://hdl.handle.net/20.500.12105/26575 Virus neutralisation assay Echovirus Enterovirus Standardisation Surveillance Assessment of twelve echovirus virus-neutralisation assays in Europe: recommendations for harmonisation of non-polio enterovirus sero-surveillance studies