Inter-Laboratory Validation of Nodal/Paranodal Antibody Testing
Por:
Lleixà, C, Titulaer, M, Rohrbacher, S, Mgbachi, V, Halstead, S, Fehmi, J, Pascual-Goñi, E, Zhu, L, Appeltshauser, L, Franken, S, Paunovic, M, Waters, P, Willison, H, Sommer, C, Querol, L, Huizinga, R, Doppler, K, Rinaldi, S
Publicada:
29 ene 2025
Resumen:
Background and AimsReliable detection of antibodies against nodal targets is vital for the diagnosis of autoimmune nodopathies. The performance characteristics of recently developed in-house assays are unknown. We compared testing at four centres.MethodsEach submitted 29-40 serum samples to a coordinating centre from one of three groups: (1) autoimmune nodopathy patients, with positive nodal/paranodal antibodies; (2) seronegative patients with other inflammatory neuropathies, and (3) healthy individuals or those with other neurological diseases. The coordinating centre recoded all samples and returned 160 identical aliquots to each testing centre for blinded testing. Once data from all centres had been received by the coordinating centre, unblinded results were returned for analysis. Sensitivity was defined by the proportion of group 1 samples returned as positive. Accuracy was defined as 0.075(sensitivity) + 0.925(specificity).ResultsCentres performed various combinations of ELISA, cell-based (CBAs) and teased-nerve fibre assays. All labs produced highly accurate results (96%-100%) and concordance for the overall result across at least 3 or all 4 test centres was observed for 98% and 89% of the samples respectively. However, 10/30 individual assays (6/14 CBAs and 4/16 ELISAs) were less than 90% sensitive. Only 3 assays had more than 1 false positive result (2 ELISAs and 1 CBA). Combining different assay modalities to produce an overall result did not improve accuracy. Inter-laboratory consistency in the determination of antibody subclasses was poor.InterpretationAlthough most samples were correctly categorised in all 4 centres, the use of a specific test modality or multiple tests did not guarantee accuracy. Early and repeated interlaboratory testing with sharing of samples is important to understand test performance and reproducibility, identify areas for improvement and maintain consistency. To aid this, we provide detailed methods for the best performing tests. Further standardisation of antibody subclass determination is required.
Filiaciones:
Lleixà, C:
Hosp Sant Creu i Sant Pau, Neuromuscular Dis Unit, Barcelona, Spain
Ctr Invest Red Enfermedades Raras CIBERER, Madrid, Spain
Titulaer, M:
Erasmus MC, Univ Med Ctr, Dept Neurol, Rotterdam, Netherlands
Rohrbacher, S:
Univ Hosp Wurzburg, Dept Neurol, Wurzburg, Germany
Mgbachi, V:
Univ Oxford, Nuffield Dept Clin Neurosci, Oxford, England
Halstead, S:
Univ Glasgow, Sch Infect & Immun, Glasgow, Scotland
Fehmi, J:
Univ Oxford, Nuffield Dept Clin Neurosci, Oxford, England
Pascual-Goñi, E:
Hosp Sant Creu i Sant Pau, Neuromuscular Dis Unit, Barcelona, Spain
Ctr Invest Red Enfermedades Raras CIBERER, Madrid, Spain
Zhu, L:
Erasmus MC, Univ Med Ctr, Dept Immunol, Rotterdam, Netherlands
Appeltshauser, L:
Univ Hosp Wurzburg, Dept Neurol, Wurzburg, Germany
Franken, S:
Erasmus MC, Univ Med Ctr, Dept Neurol, Rotterdam, Netherlands
Paunovic, M:
Erasmus MC, Univ Med Ctr, Dept Neurol, Rotterdam, Netherlands
Waters, P:
Univ Oxford, Nuffield Dept Clin Neurosci, Oxford, England
Willison, H:
Univ Glasgow, Sch Infect & Immun, Glasgow, Scotland
Sommer, C:
Univ Hosp Wurzburg, Dept Neurol, Wurzburg, Germany
Querol, L:
Hosp Sant Creu i Sant Pau, Neuromuscular Dis Unit, Barcelona, Spain
Ctr Invest Red Enfermedades Raras CIBERER, Madrid, Spain
Huizinga, R:
Erasmus MC, Univ Med Ctr, Dept Immunol, Rotterdam, Netherlands
Doppler, K:
Univ Hosp Wurzburg, Dept Neurol, Wurzburg, Germany
Rinaldi, S:
Univ Oxford, Nuffield Dept Clin Neurosci, Oxford, England
Green Submitted, Green Accepted, hybrid
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