Caudie Christiane, Quittard Pinon Arnaud, Bouhour Françoise, Vial Christophe, Garnier Lorna, Fabien Nicole
Hôpitaux de Lyon Laboratoire d'Immunologie, Groupement Hospitalier Lyon Sud 69495 Pierre-Bénite, France.
Hôpitaux de Lyon Service Electroneuromyographie et Pathologies Neuromusculaires, Groupement Hospitalier Est, Hôpital Neurologique, 69677 Lyon Bron, France.
Clin Lab. 2013;59(11-12):1277-87. doi: 10.7754/clin.lab.2013.121116.
To assess the performance of commercial anti-ganglioside antibody assays, we determined anti-ganglioside antibody IgG and IgM isotype profiles of patients with acute and chronic well-characterized immune-mediated peripheral neuropathies by one immunodot assays (Zentec/Ingen: Dotzen Ganglio Profile Ab, Euroimmun/BioAdvance: Euroline ganglioprofile), two line-immuno assay (GA Generic Assays/Labodia: Anti-Gangli osid Dot, Euroimmun/BioAdvance: Euroline ganglioprofile), and one enzyme-linked immunosorbent assay (ELISA) (Bühlmann: GanglioCombi). Specific antibody profiles were compared with those obtained by our validated standard in-house immunodot assay (IDA).
We selected 33 sera with high levels of IgG and IgM anti-ganglioside antibodies from 15 patients with Guillain-Barre syndrome (GBS) subtypes and variants, 12 patients with CANOMAD syndrome (chronic ataxic neuropathy with ophthalmoplegia, M-paraprotein, cold agglutinins, disialosyl antibodies), 5 patients with chronic motor peripheral neuropathies, and 1 patient with sensory neuropathy and a control group composed of 10 patients with non-autoimmune neuropathy.
The 3 commercial IDAs employing hydrophobic membranes and the ELISA demonstrated different carbohydrate epitopes on 6 to 12 glycolipid antigens used for anti-ganglioside antibody detection. Comparison with the validated in-house IDA showed large variations in sensitivity between tests and a more diverse reactivity to gangliosides than expected. The test with the largest panel of glycolipids detecting 11 anti-ganglioside antibody reactivities (GM1, GM2, GM3, GM4, GD1a, GD1b, GD2, GD3, GT1a, GT1b, GQ1b, and sulfatide) revealed the best concordance with our in-house assay. However, even with this test, differences were observed in the immunoreactivity against some gangliosides and weakly stained bands were not easy to interpret.
Our data suggest an urgent need for standardization of commercial anti-ganglioside assays and the introduction of international anti-ganglioside antibody reference standards.
为评估商用抗神经节苷脂抗体检测方法的性能，我们通过一种免疫斑点法（Zentec/Ingen：Dotzen Ganglio Profile Ab，Euroimmun/BioAdvance：Euroline ganglioprofile）、两种线性免疫分析法（GA Generic Assays/Labodia：Anti-Gangli osid Dot，Euroimmun/BioAdvance：Euroline ganglioprofile）和一种酶联免疫吸附测定法（ELISA）（Bühlmann：GanglioCombi），测定了急性和慢性特征明确的免疫介导性周围神经病患者的抗神经节苷脂抗体IgG和IgM同种型谱。将特异性抗体谱与我们经验证的标准内部免疫斑点法（IDA）所获得的谱进行比较。
我们从15例吉兰 - 巴雷综合征（GBS）亚型和变异型患者、12例CANOMAD综合征（伴有眼肌麻痹、M蛋白、冷凝集素、二唾液酸抗体的慢性共济失调性神经病）患者、5例慢性运动性周围神经病患者和1例感觉神经病患者中选取了33份抗神经节苷脂抗体IgG和IgM水平较高的血清，并设立了一个由10例非自身免疫性神经病患者组成的对照组。
3种采用疏水膜的商用免疫斑点法和ELISA在用于抗神经节苷脂抗体检测的6至12种糖脂抗原上显示出不同的碳水化合物表位。与经验证的内部IDA相比，各检测方法之间的敏感性差异很大，且对神经节苷脂的反应性比预期更为多样。检测糖脂种类最多、能检测11种抗神经节苷脂抗体反应性（GM1、GM2、GM3、GM4、GD1a、GD1b、GD2、GD3、GT1a、GT1b、GQ1b和硫苷脂）的检测方法与我们的内部检测方法一致性最佳。然而，即便使用这种检测方法，在针对某些神经节苷脂的免疫反应性方面仍存在差异，且弱阳性条带不易解读。
我们的数据表明，迫切需要对商用抗神经节苷脂检测方法进行标准化，并引入国际抗神经节苷脂抗体参考标准。
