Detection of Autoantibodies Against the Acetylcholine Receptor, Evaluation of Commercially Available Methodologies: Fixed Cell-Based Assay, Radioimmunoprecipitation Assay and Enzyme-Linked Immunosorbent Assay1.

IF 4.3 3区材料科学 Q1 ENGINEERING, ELECTRICAL & ELECTRONIC

ACS Applied Electronic Materials Pub Date : 2024-01-01 DOI:10.3233/JND-230210

Larissa Diogenes, Alessandra Dellavance, Danielle Cristiane Baldo, Sarah Cristina Gozzi-Silva, Kethellen Gomes, Monica Simon Prado, Luis Eduardo C Andrade, Gerson Dierley Keppeke

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引用次数: 0

Abstract

Background/objective: Myasthenia Gravis (MG) is an autoimmune disorder characterized by pathogenic autoantibodies (AAbs) targeting nicotinic acetylcholine receptors (AChR), disrupting neuromuscular communication. RadioImmunoPrecipitation Assay (RIPA) is recommended to detect AChR AAbs, but its complexity and radioactive requirements limit widespread use. We compare non-RIPA anti-AChR immunoassays, including Cell-Based Assay (CBA) and two ELISA kits, against the gold standard RIPA.

Methods/results: 145 samples were included with medical indication for anti-AChR testing. By the RIPA method, 63 were negative (RIPA-Neg < 0.02 nmol/L), 18 were classified as Borderline (≥0.02 -1 nmol/L), and 64 were positive (RIPA-Pos > 1 nmol/L). The competitive ELISA showed poor agreement with RIPA (Kappa = 0.216). The indirect ELISA demonstrated substantial agreement with RIPA (Kappa = 0.652), with ∼76% sensitivity and ∼94% specificity for MG diagnostic. The CBA, where fixed cells expressing clustered AChR were used as substrate, exhibited almost perfect agreement with RIPA (Kappa = 0.984), yielding ∼98% sensitivity and 96% specificity for MG. In addition, a semiquantitative analysis showed a strong correlation between CBA titration, indirect ELISA, and RIPA levels (r = 0.793 and r = 0.789, respectively).

Conclusions: The CBA displayed excellent analytical performance for MG diagnostic when compared to RIPA, making it a potential replacement for RIPA in clinical laboratories. Some solid-phase assays (such as the indirect ELISA applied here), as well as CBA titration, offer reliable options to estimate anti-AChR AAb levels after confirming positivity by the CBA.∥.

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检测乙酰胆碱受体自身抗体，评估市售方法：固定细胞测定法、放射免疫沉淀测定法和酶联免疫吸附测定法1。

背景/目的：重症肌无力（MG）是一种自身免疫性疾病，其特征是以烟碱乙酰胆碱受体（AChR）为靶点的致病性自身抗体（AAbs）会破坏神经肌肉的通讯。放射免疫沉淀法（RIPA）被推荐用于检测 AChR AAbs，但其复杂性和放射性要求限制了其广泛应用。我们将非 RIPA 抗 AChR 免疫分析法（包括细胞分析法（CBA）和两种 ELISA 试剂盒）与黄金标准 RIPA 进行了比较：方法/结果：145 份样本具有抗 AChR 检测的医学指征。用 RIPA 方法检测，63 份呈阴性（RIPA-Neg 1 nmol/L）。竞争性 ELISA 与 RIPA 的一致性较差（Kappa = 0.216）。间接 ELISA 与 RIPA 的吻合度很高（Kappa = 0.652），对 MG 诊断的敏感性和特异性分别为 76% 和 94%。以表达簇状 AChR 的固定细胞为底物的 CBA 与 RIPA 几乎完全一致（Kappa = 0.984），对 MG 的敏感性和特异性分别为 98% 和 96%。此外，半定量分析显示 CBA 滴定、间接 ELISA 和 RIPA 水平之间存在很强的相关性（r = 0.793 和 r = 0.789）：结论：与 RIPA 相比，CBA 在 MG 诊断方面显示出卓越的分析性能，使其成为临床实验室中 RIPA 的潜在替代品。一些固相检测方法（如本文应用的间接ELISA）以及CBA滴定法为CBA确认阳性后估算抗ACHR AAb水平提供了可靠的选择。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

ACS Applied Electronic Materials Multiple-

CiteScore

7.20

自引率

4.30%

发文量

567