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....

Celý popis

Uloženo v:

Podrobná bibliografie
Vydáno v:	PloS one Ročník 10; číslo 3; s. e0122037
Hlavní autoři:	Ramberger, Melanie, Peschl, Patrick, Schanda, Kathrin, Irschick, Regina, Höftberger, Romana, Deisenhammer, Florian, Rostásy, Kevin, Berger, Thomas, Dalmau, Josep, Reindl, Markus
Médium:	Journal Article
Jazyk:	angličtina
Vydáno:	United States Public Library of Science 27.03.2015 Public Library of Science (PLoS)
Témata:	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
On-line přístup:	Získat plný text
Tagy:	Přidat tag Žádné tagy, Buďte první, kdo vytvoří štítek k tomuto záznamu!

Popis
Shrnutí:	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.
Bibliografie:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 14 ObjectType-Article-2 ObjectType-Feature-1 content type line 23 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