PMID- 25815887
OWN - NLM
STAT- MEDLINE
DCOM- 20160223
LR  - 20220129
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 10
IP  - 3
DP  - 2015
TI  - 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.
PG  - e0122037
LID - 10.1371/journal.pone.0122037 [doi]
LID - e0122037
AB  - 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.
FAU - Ramberger, Melanie
AU  - Ramberger M
AD  - Clinical Department of Neurology, Medical University of Innsbruck, Innsbruck, 
      Austria.
FAU - Peschl, Patrick
AU  - Peschl P
AD  - Clinical Department of Neurology, Medical University of Innsbruck, Innsbruck, 
      Austria.
FAU - Schanda, Kathrin
AU  - Schanda K
AD  - Clinical Department of Neurology, Medical University of Innsbruck, Innsbruck, 
      Austria.
FAU - Irschick, Regina
AU  - Irschick R
AD  - Division of Neuroanatomy, Medical University of Innsbruck, Innsbruck, Austria.
FAU - Hoftberger, Romana
AU  - Hoftberger R
AD  - Institute of Neurology, Medical University of Vienna, Vienna, Austria; Institucio 
      Catalana de Recerca i Estudis Avancats (ICREA), IDIBAPS, Hospital Clinic, 
      Barcelona, Spain.
FAU - Deisenhammer, Florian
AU  - Deisenhammer F
AD  - Clinical Department of Neurology, Medical University of Innsbruck, Innsbruck, 
      Austria.
FAU - Rostasy, Kevin
AU  - Rostasy K
AD  - Pediatric Neurology, Witten/Herdecke University, Children's Hospital Datteln, 
      Datteln, Germany.
FAU - Berger, Thomas
AU  - Berger T
AD  - Clinical Department of Neurology, Medical University of Innsbruck, Innsbruck, 
      Austria.
FAU - Dalmau, Josep
AU  - Dalmau J
AD  - Institucio Catalana de Recerca i Estudis Avancats (ICREA), IDIBAPS, Hospital 
      Clinic, Barcelona, Spain.
FAU - Reindl, Markus
AU  - Reindl M
AD  - Clinical Department of Neurology, Medical University of Innsbruck, Innsbruck, 
      Austria.
LA  - eng
GR  - R01 NS077851/NS/NINDS NIH HHS/United States
GR  - W 1206/FWF_/Austrian Science Fund FWF/Austria
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20150327
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (Autoantibodies)
RN  - 0 (Receptors, N-Methyl-D-Aspartate)
SB  - IM
MH  - Adolescent
MH  - Adult
MH  - Anti-N-Methyl-D-Aspartate Receptor Encephalitis/*blood
MH  - Autoantibodies/*blood
MH  - Case-Control Studies
MH  - Child
MH  - Female
MH  - Flow Cytometry/methods
MH  - HEK293 Cells
MH  - Humans
MH  - Male
MH  - Microscopy, Fluorescence/methods
MH  - Receptors, N-Methyl-D-Aspartate/immunology
MH  - Sensitivity and Specificity
MH  - Serologic Tests/*methods
PMC - PMC4376531
COIS- 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.
EDAT- 2015/03/31 06:00
MHDA- 2016/02/26 06:00
PMCR- 2015/03/27
CRDT- 2015/03/28 06:00
PHST- 2014/07/08 00:00 [received]
PHST- 2015/02/06 00:00 [accepted]
PHST- 2015/03/28 06:00 [entrez]
PHST- 2015/03/31 06:00 [pubmed]
PHST- 2016/02/26 06:00 [medline]
PHST- 2015/03/27 00:00 [pmc-release]
AID - PONE-D-14-30477 [pii]
AID - 10.1371/journal.pone.0122037 [doi]
PST - epublish
SO  - PLoS One. 2015 Mar 27;10(3):e0122037. doi: 10.1371/journal.pone.0122037. 
      eCollection 2015.