Comparison of Diagnostic Accuracy of Microscopy and Flow Cytometry in Evaluating N-Methyl-D-Aspartate Receptor Antibodies in Serum Using a Live Cell-Based Assay

N-methyl-D-aspartate receptor (NMDAR) encephalitis is an autoimmune neurological disease, diagnosed by a specific autoantibody against NMDAR. Antibody testing using commercially available cell-based assays (CBA) or immunohistochemistry on rat brain tissue has proven high specificity and sensitivity....

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Bibliographic Details
Published in:PloS one Vol. 10; no. 3; p. e0122037
Main Authors: Ramberger, Melanie, Peschl, Patrick, Schanda, Kathrin, Irschick, Regina, Höftberger, Romana, Deisenhammer, Florian, Rostásy, Kevin, Berger, Thomas, Dalmau, Josep, Reindl, Markus
Format: Journal Article
Language:English
Published: United States Public Library of Science 27.03.2015
Public Library of Science (PLoS)
Subjects:
Adolescent
Adult
Analysis
Animal tissues
Anti-N-Methyl-D-Aspartate Receptor Encephalitis - blood
Antibodies
Antigens
Aspartate
Assaying
Autoantibodies
Autoantibodies - blood
Autoimmune diseases
Autoimmunity
Brain
Case-Control Studies
Child
Confidence intervals
Diagnostic systems
Disease
Encephalitis
Female
Flow cytometry
Flow Cytometry - methods
Fluorescence
Glutamate receptors
Glutamic acid receptors
HEK293 Cells
Hospitals
Humans
Immunofluorescence
Immunoglobulins
Immunohistochemistry
Laboratories
Male
Medical diagnosis
Microscopy
Microscopy, Fluorescence - methods
Multiple sclerosis
N-methyl-D-aspartate
N-Methyl-D-aspartic acid receptors
Neurological diseases
Neurology
Patients
Performance evaluation
Receptors, N-Methyl-D-Aspartate - immunology
Rodents
Sensitivity
Sensitivity analysis
Sensitivity and Specificity
Serologic Tests - methods
Signal transduction
Austria
Germany
Spain
ISSN:1932-6203, 1932-6203
Online Access:Get full text
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Description
Summary:N-methyl-D-aspartate receptor (NMDAR) encephalitis is an autoimmune neurological disease, diagnosed by a specific autoantibody against NMDAR. Antibody testing using commercially available cell-based assays (CBA) or immunohistochemistry on rat brain tissue has proven high specificity and sensitivity. Here we compare an immunofluorescence live CBA to a flow cytometry (FACS) based assay to detect NMDAR antibodies by their binding to the surface of HEK293A cells functionally expressing NMDAR. Both assays were first established using a discovery group of 76 individuals and then validated in a group of 32 patients in a blinded manner. In the CBA, 23 of 23 patients with NMDAR encephalitis were positive for NMDAR antibodies and 0 of 85 controls (32 healthy controls and 53 patients with other neurological diseases), resulting in a sensitivity and specificity of 100% (95% confidence intervals (CI) 85.1-100.0 and 95.7-100.0, respectively). The FACS based assay detected NMDAR antibodies in 20 of 23 patients and in 0 of 85 controls. Therefore, with an equally high specificity (95% CI 95.7-100.0) the sensitivity of the FACS based assay was 87% (95% CI 66.4-97.2). Comparing antibody titers from CBA with delta median fluorescence intensities from FACS showed a high concordance (kappa = 0.943, p<0.0001) and correlation (r = 0.697, p<0.0001). In conclusion, evaluation of the FACS based assay revealed a lower sensitivity and high inter-assay variation, making the CBA a more reliable detection method.
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Conceived and designed the experiments: M. Ramberger M. Reindl. Performed the experiments: M. Ramberger PP KS RI M. Reindl. Analyzed the data: M. Ramberger M. Reindl. Contributed reagents/materials/analysis tools: RH FD KR TB JD. Wrote the paper: M. Ramberger RH JD M. Reindl.
Competing Interests: JD has received a research grant from Euroimmun and holds a patent (Methods and compositions for treatment and diagnosis of encephalitis or epilepsy, USA patent number 7,972,796 B2, Europe EP 2 057 466) for the use of NMDA receptor as autoantibody test; he receives royalties for this antibody testing. RH and FD offer commercial testing for NMDA receptor antibodies. Markus Reindl is a PLOS ONE Editorial Board member. This does not alter the authors' adherence to PLOS ONE Editorial policies and criteria.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0122037