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

Autor(es)

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

• Tipo de documento: Artigo Científico
• Fonte: JOURNAL OF NEUROMUSCULAR DISEASES; v. 11, n. 3, p. 11-pg., 2024-01-01.

Resumo

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 detectAChR 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 similar to 76% sensitivity and similar to 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 similar to 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. (AU)

• Processo FAPESP: 21/04588-9 - Construção de um ensaio baseado em células para detecção de anticorpos contra o receptor de acetilcolina
• Beneficiário: Larissa Diogenes Santos
• Modalidade de apoio: Bolsas no Brasil - Mestrado
• Processo FAPESP: 17/20745-1 - Depleção de autoanticorpos por edição gênica com o sistema CRISPR/Cas9 em plasmócitos autorreativos
• Beneficiário: Gerson Dierley Keppeke
• Modalidade de apoio: Auxílio à Pesquisa - Jovens Pesquisadores