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High-throughput Flow Cytometry Cell-based Assay to Detect Antibodies to N-Methyl-D-aspartate Receptor or Dopamine-2 Receptor in Human Serum
Published on: November 23, 2013
TE671 cell-based ELISA for anti-acetylcholine receptor antibody determination in myasthenia gravis
D Franciotta1, G Martino, E Brambilla
1Laboratory of Neuroimmunology, Istituto di Ricovera e Cura a Carattere Scientifico, Neurological Institute Mondino, University of Pavia, Italy. francid@cpbim1.unipv.it
Background:
Acetylcholine receptor (AChR) from human muscles is the antigen used currently in radioimmunoprecipitation assays (RIPAs) for the determination of anti-AChR antibodies in the diagnosis of myasthenia gravis (MG). Our aim was to develop and validate an ELISA using TE671 cells as the source of AChR.
Methods:
After TE671 cell homogenization, the crude AChR extract was used for plate coating. Anti-AChR antibodies were determined in 207 MG patients and in 77 controls.
Results:
The mean intra- and interassay CVs (for two samples with different anti-AChR antibody concentrations) were 9.7% and 15.7%, respectively. Test sensitivity and specificity, for generalized MG, were 79.5% (95% confidence interval, 72.8-85.0%) and 96.1% (89.0-99.1%). The detection limit was 2 nmol/L. Anti-AChR antibody concentrations from 53 MG patients, as tested with our ELISA, showed good agreement with an RIPA with a mean difference (SD) of 1.0 (5.6) nmol/L.
Conclusion:
Our ELISA is a simple screening test for the diagnosis of MG and enables rapid and inexpensive patient follow-up.
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