Doğan İpek Güngör, Yiğit Mesut, Tezer Damla Çetinkaya, Gülaçtı Özlem, Ayhan Şevval, Türkdemir İpek Duygu, Çelik Betül, Taşdelen Beril, Uzunköprü Cihat, Yetkin Mehmet Fatih, Tütüncü Melih, Kilercik Meltem, Demir Serkan
Department of Neurology, University of Health Sciences, Sancaktepe Sehit Prof Dr Ilhan Varank Training and Research Hospital, Istanbul, Türkiye.
Acibadem Mehmet Ali Aydinlar University, Human Immunology Center, Istanbul, Türkiye.
Noro Psikiyatr Ars. 2025 Jul 29;62(3):259-263. doi: 10.29399/npa.28951. eCollection 2025.
Neuromyelitis Optica (NMO) is an inflammatory disorder affecting the central nervous system, notably the optic nerve and spinal cord. Seropositive NMO is marked by serum IgG antibodies against aquaporin-4 (AQP4). The accurate identification of AQP4-IgG is crucial for distinguishing NMO from other demyelinating diseases of the central nervous system. However, traditional diagnostic assays have limitations in sensitivity and specificity. Here, we introduce our in-house flow cytometry live cell-based assay (FC-LCBA) for detecting AQP4 antibodies with enhanced sensitivity and specificity. Our objective is to report the accuracy and compare the efficacy of our newly developed in-house FC-LCBA against the commercial cell-based indirect immunofluorescence assay (IIFA) in detecting AQP4 antibodies.
This single-blind study was approved by the ethical committee and involved 101 serum samples. Twenty-five samples (including retests) from 17 patients evaluated in the NMO spectrum who had at least one positive cell-based IIFA test during the diagnosis or follow-up are tested in parallel with our in-house FC-LCBA and cell-based IIFA. In addition, 36 serum samples from myelin oligodendrocyte glycoprotein-associated disease (MOGAD) patients and 40 serum samples from healthy subjects are also referred for specificity analysis.
Our in-house FC-LCBA displayed superior sensitivity, detecting positive results even when the cell-based IIFA yielded negative results in patients under immunosuppressive treatments. Additionally, FC-LCBA exhibited high specificity for NMO, showing negligible antibody levels in patients with MOGAD diagnosis and healthy individuals. The assay's stability was confirmed through consistent results in retests.
Our in-house FC-LCBA emerges as a promising diagnostic tool for detecting AQP4 antibodies, offering improved sensitivity, specificity, and reliability, instilling confidence in its potential.
视神经脊髓炎（NMO）是一种影响中枢神经系统的炎症性疾病，尤其是视神经和脊髓。血清阳性的NMO以抗水通道蛋白4（AQP4）的血清IgG抗体为特征。准确识别AQP4-IgG对于区分NMO与其他中枢神经系统脱髓鞘疾病至关重要。然而，传统诊断方法在敏感性和特异性方面存在局限性。在此，我们介绍我们基于流式细胞术活细胞检测法（FC-LCBA），用于检测AQP4抗体，具有更高的敏感性和特异性。我们的目的是报告我们新开发的内部FC-LCBA在检测AQP4抗体方面的准确性，并与基于细胞的商业间接免疫荧光法（IIFA）的效能进行比较。
这项单盲研究经伦理委员会批准，涉及101份血清样本。从17例在NMO谱系中评估的患者中选取25份样本（包括复测样本），这些患者在诊断或随访期间至少有一次基于细胞的IIFA检测呈阳性，将其与我们的内部FC-LCBA和基于细胞的IIFA同时进行检测。此外，还送检了36份来自髓鞘少突胶质细胞糖蛋白相关疾病（MOGAD）患者的血清样本和40份来自健康受试者的血清样本进行特异性分析。
我们的内部FC-LCBA显示出更高的敏感性，即使在免疫抑制治疗的患者中，当基于细胞的IIFA检测结果为阴性时，它也能检测到阳性结果。此外，FC-LCBA对NMO表现出高特异性，在诊断为MOGAD的患者和健康个体中抗体水平可忽略不计。通过复测结果的一致性证实了该检测方法的稳定性。
我们的内部FC-LCBA成为一种有前景的检测AQP4抗体的诊断工具，具有更高的敏感性、特异性和可靠性，让人对其潜力充满信心。
