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, Kimberley S.M. Benschop3, Gerrit Koen2, Hetty van Eijk2, Heli Harvala4,5, Jean-Luc Bailly6, Maël Bessaud7, Everlyn Kamau8, Isabelle Simon6, Marie-Line Joffret7, Lubomira Nikolaeva-Glomb9, Irina Georgieva9, Asya Stoyanova9, Sabine Diedrich10, Sindy Böttcher10, Maria Cabrerizo11, Irena Tabain12, Željka Hruškar12, Vladimir Stevanović13, Petri Susi14, Eero Hietanen14,15, Paula Palminha16, Petra Rainetová17, Anda Baicus18, Maartje Kristensen1, Dasja Pajkrt19, Jean-Luc Murk1, Katja C. Wolthers2 and European non-polio enterovirus network20
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.
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