Diagnostic sensitivity, specificity and likelihood ratios (LR+, LR−) for NMO calculated for each single substrate of the BioChip, obtained by testing 91 serum samples on the “Neurology Mosaic 17”.

<p>NMO: neuromyelitis optica, LR: likelihood ratio.</p

Association between fluorescence patterns and clinical status of all the analyzed samples (2 commercial anti-AQP4 positive controls, 20 NMO samples, 41 MS samples, 30 healthy subjects).

<p>NMO: neuromyelitis optica, MS: multiple sclerosis, HC: healthy controls.</p

Differential distribution of IIF staining patterns in five diagnostic substrates following incubation with serum samples from patients with NMO or controls.

<p>The corresponding staining patterns (A, B, C, D; typical and atypical white matter staining), as defined in the results section, are indicated, together with the final evaluation of positivity or negativity for anti-AQP4 antibodies, and the healthy volunteers' clinical status. NMO: neuromyelitis optica, MS: multiple sclerosis, HC: healthy controls, CNT 1: commercial goat polyclonal anti-human AQP4 IgG (H19, Santa Cruz Biotechnology), CNT 2: human anti-AQP4 positive serum provided by the manufacturer, CNT 3: human anti-AQP4 negative serum provided by the manufacturer, TC = transfected cells, NTC = non-transfected cells, ON = optic nerve.</p

“Typical” (A) and “atypical” (B) white matter fluorescence staining as observed with anti-AQP4 positive NMO (A) and anti-AQP4 negative MS (B) human serum samples, respectively.

<p>Bound IgG was visualized using secondary antibodies labeled with FITC (green). Cell nuclei, stained with DAPI, are shown in blue. Magnification 40×. NMO: neuromyelitis optica; MS: multiple sclerosis; DAPI: 4,6-diamidino-2-phenylindole; FITC: Fluorescein isothiocyanate.</p

Demographic and clinical characteristics of subjects.

<p>NMO: neuromyelitis optica, MS: multiple sclerosis, HC: healthy controls, MRI: magnetic resonance imaging, CSF: cerebrospinal fluid, OB: oligoclonal bands.</p>*<p>One NMO patient and one MS patient are pediatric.</p>**<p>According to Wingerchuk 2006, where MRI Paty criteria for MS were included.</p>***<p>CSF data were not available for 2 NMO patients and for 4 MS patients.</p

Fluorescence staining patterns as observed with positive and negative controls provided by the manufacturer (column 1 and 2, respectively), an anti-AQP4 antibody positive human serum sample (column 3), and an anti-AQP4 antibody negative human control sample (column 4).

<p>The biochip mosaic consists of 5 substrates: HEK cells transfected with full length recombinant human AQP4 (row A), non-transfected HEK cells (row B) and cryosections of primate cerebellum (row C), cerebrum (row D) and optic nerve (row E). Bound IgG was visualized using secondary antibodies labeled with FITC (green). Cell nuclei, stained with DAPI, are shown in blue. Magnification 40×. GL: granular layer; WM: white matter; DAPI: 4,6-diamidino-2-phenylindole; FITC: fluorescein isothiocyanate.</p