ELISA and MAP-PCR results.

<p>Antibody positive patients were so identified by selecting a cut-off OD value of 0.84 (as calculated by the mean plus two times the standard deviation of the mean).</p

Blast alignments with masking-off segments of the query sequence that have low compositional complexity.

<p>Results show region of identity, strongly similar amino acids with a plus (+) and missing region in dashes (–). Query sequence is MAP2694.</p

AUC-ROC Test.

<p>Evaluation of serum samples from 50 MS patients (right column) and 56 healthy controls (left column) against MAP2694 recombinant protein by the AUC-ROC test. The horizontal blue line highlights the cut-off value(s) which gives a specificity of 25%. Data are presented as OD values observed in ELISA. Area under curve (AUC) = 0,686; P = 0,0003.</p

Evaluation of serum samples against MAP2694 recombinant protein.

<p>MS patients sera are shown in the left column whereas Healthy Controls sera are shown in the right column. Data are presented as OD values observed in ELISA, as also described in the text. Data from a representative experiment out of three are shown. The mean value for each group is indicated by a thick horizontal line. IS900 PCR positive samples are indicated as black solid figures.</p

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

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

“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