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Assessment of Twelve Echovirus Virus-Neutralisation Assays in Europe: Recommendations for Harmonisation of Non-Polio Enterovirus Sero-Surveillance Studies

JOURNAL OF GENERAL VIROLOGY(2024)

Elisabeth TweeSteden Hosp | Natl Inst Publ Hlth & Environm | Amsterdam UMC | NHS Blood & Transplant | Univ Clermont Auvergne | Univ Oxford | Natl Ctr Infect & Parasit Dis | Robert Koch Inst | Croatian Inst Publ Hlth | Croatian Institute of Public Health | Fac Vet Med | Univ Turku | Natl Inst Hlth | Natl Inst Publ Hlth NRL Enteroviruses | Cantacuzino Natl Inst Med Mil Res & Dev

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Abstract
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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Virus neutralisation assay,enterovirus,echovirus,surveillance,standardisation
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要点】:本文评估了欧洲十二个实验室的非脊髓灰质炎肠道病毒中和试验,发现实验方法差异较大,建议进行标准化以改进病毒中和抗体滴度的准确性。

方法】:通过欧洲非脊髓灰质炎肠道病毒网络,对参与实验室提供的五种肠道病毒类型(E1、E18、E30 G2、E30 G6和E6)及静脉免疫球蛋白(IVIG)样本进行病毒中和试验,分析各实验室的实验方法和抗体滴度。

实验】:十二个实验室参与的质量评估实验中,发现病毒中和试验的细胞系类型对结果有显著影响,但标准化细胞系传代数并未显著减少实验室间的变异;使用参考样本进行校准可减少变异,但抗体滴度差异仍大于两个log2稀释步骤。