Lleixà Cinta, Titulaer Maarten, Rohrbacher Sophia, Mgbachi Victor, Halstead Susan, Fehmi Janev, Pascual-Goñi Elba, Zhu Louisa, Appeltshauser Luise, Franken Suzanne, Paunovic Manuela, Waters Patrick, Willison Hugh, Sommer Claudia, Querol Luis, Huizinga Ruth, Doppler Kathrin, Rinaldi Simon
Neuromuscular Diseases Unit, Hospital de la Santa Creu i Sant Pau, Barcelona, Spain.
Centro Para la Investigación en Red en Enfermedades Raras (CIBERER), Madrid, Spain.
J Peripher Nerv Syst. 2025 Mar;30(1):e70000. doi: 10.1111/jns.70000.
Reliable 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.
Each 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).
Centres 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.
Although 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.
可靠检测针对神经节靶点的抗体对于自身免疫性神经节病的诊断至关重要。目前尚不清楚最近开发的内部检测方法的性能特征。我们比较了四个中心的检测情况。
每个中心从三组中的一组向协调中心提交29 - 40份血清样本：（1）自身免疫性神经节病患者，其神经节/神经旁抗体呈阳性；（2）血清阴性的其他炎性神经病患者；（3）健康个体或患有其他神经系统疾病的个体。协调中心对所有样本进行重新编码，并将160份相同的样本等分送回每个检测中心进行盲法检测。一旦协调中心收到所有中心的数据，便会将未盲法的结果返回进行分析。敏感性由第1组样本中呈阳性返回的比例定义。准确性定义为0.075（敏感性）+ 0.925（特异性）。
各中心采用了酶联免疫吸附测定（ELISA）、基于细胞的检测方法（CBA）和 teased - 神经纤维检测等多种组合。所有实验室均得出了高度准确的结果（96% - 100%），分别有98%和89%的样本在至少3个或所有4个检测中心观察到总体结果的一致性。然而，30项单独检测中有10项（14项CBA中的6项和16项ELISA中的4项）敏感性低于90%。只有3项检测有超过1例假阳性结果（2项ELISA和1项CBA）。组合不同的检测方法得出总体结果并未提高准确性。实验室间在抗体亚类测定方面的一致性较差。
尽管所有4个中心对大多数样本都进行了正确分类，但使用特定的检测方法或多种检测并不能保证准确性。早期和反复的实验室间检测并共享样本对于了解检测性能和可重复性、确定改进领域以及保持一致性很重要。为了便于此操作，我们提供了性能最佳检测的详细方法。抗体亚类测定需要进一步标准化。
