Serologic diagnosis of NMO: a multicenter comparison of aquaporin-4-IgG assays.
OBJECTIVE
Neuromyelitis optica (NMO) immunoglobulin G (IgG) (aquaporin-4 [AQP4] IgG) is highly specific for NMO and related disorders, and autoantibody detection has become an essential investigation in patients with demyelinating disease. However, although different techniques are now used, no multicenter comparisons have been performed. This study compares the sensitivity and specificity of different assays, including an in-house flow cytometric assay and 2 commercial assays (ELISA and transfected cell-based assay [CBA]).


METHODS
Six assay methods (in-house or commercial) were performed in 2 international centers using coded serum from patients with NMO (35 patients), NMO spectrum disorders (25 patients), relapsing-remitting multiple sclerosis (39 patients), miscellaneous autoimmune diseases (25 patients), and healthy subjects (22 subjects).


RESULTS
The highest sensitivities were yielded by assays detecting IgG binding to cells expressing recombinant AQP4 with quantitative flow cytometry (77; 46 of 60) or visual observation (CBA, 73%; 44 of 60). The fluorescence immunoprecipitation assay and tissue-based immunofluorescence assay were least sensitive (48%-53%). The CBA and ELISA commercial assays (100% specific) yielded sensitivities of 68% (41 of 60) and 60% (36 of 60), respectively, and sensitivity of 72% (43 of 60) when used in combination.


CONCLUSIONS
The greater sensitivity and excellent specificity of second-generation recombinant antigen-based assays for detection of NMO-IgG in a clinical setting should enable earlier diagnosis  of NMO spectrum disorders and prompt initiation of disease-appropriate therapies.
Neuroimmunology Group, Nuffield Department of Clinical Neurosciences, Oxford, UK. paddy.waters@imm.ox.ac.uk
P J
PJ
Waters
A
A
McKeon
M I
MI
Leite
S
S
Rajasekharan
V A
VA
Lennon
A
A
Villalobos
J
J
Palace
J N
JN
Mandrekar
A
A
Vincent
A
A
Bar-Or
S J
SJ
Pittock
D000328:Adult; D051401:Aquaporin 4 / Q000032:analysis* / Q000276:immunology; D006801:Humans; D007118:Immunoassay / Q000592:standards*; D007074:Immunoglobulin G / Q000032:analysis* / Q000276:immunology; D009471:Neuromyelitis Optica / Q000175:diagnosis* / Q000276:immunology; D012680:Sensitivity and Specificity
D051401:Aquaporin 4; D007074:Immunoglobulin G