Test Catalog

Test ID: NMOFS    
Neuromyelitis Optica (NMO)/Aquaporin-4-IgG Fluorescence-Activated Cell Sorting (FACS) Assay, Serum

Useful For Suggests clinical disorders or settings where the test may be helpful

Diagnosis of a neuromyelitis optica spectrum disorder (NMOSD)

 

Diagnosis of autoimmune AQP4 channelopathy

 

Diagnosis of neuromyelitis optica (NMO)

 

Distinguishing NMOSD from multiple sclerosis early in the course of disease

Testing Algorithm Delineates situations when tests are added to the initial order. This includes reflex and additional tests.

When the results of this assay require further evaluation, NMOTS / Neuromyelitis Optica (NMO)/Aquaporin-4-IgG Fluorescence-Activated Cell Sorting (FACS) Titer Assay, Serum will be performed at an additional charge.

Clinical Information Discusses physiology, pathophysiology, and general clinical aspects, as they relate to a laboratory test

Neuromyelitis optica (NMO), sometimes called Devic disease or opticospinal multiple sclerosis [MS]) is a severe, relapsing, autoimmune, inflammatory and demyelinating central nervous system disease that predominantly affects optic nerves and spinal cord.(1) The disorder is now recognized as a spectrum of autoimmunity (termed NMO spectrum disorders [NMOSD]) targeting the astrocytic water channel aquaporin-4 (AQP4).(1,2) Brain lesions are observed in >60% of patients with NMOSD and approximately 10% will be MS-like.(3) Children tend to have greater brain involvement than adults and brain lesions are more symptomatic than is typical for adult patients.(4) Extensive cerebral white matter signal abnormalities are sometimes encountered, most commonly in children, and are sometimes associated with encephalopathy. Circumventricular organs (CVO; eg, area postrema) are preferentially involved. Symptoms and signs attributable to area postrema involvement include intractable hiccups, nausea and vomiting, and these may occur in isolation, herald the onset of NMO or occur in association with the more classical optic neuritis or Longitudinally Extensive Transverse Myelitis (LETM).(5) Magnetic resonance imaging typically reveals large inflammatory spinal cord lesions involving 3 or more vertebral segments. During acute attacks, the cerebrospinal fluid contains inflammatory cells, but usually lacks evidence of intrathecal IgG synthesis. The clinical course is characterized by relapses of optic neuritis or transverse myelitis, or both. Many patients with NMOSD are misdiagnosed as having MS. Importantly, the prognosis and optimal treatments for the 2 diseases differ. NMOSD typically has a worse natural history than MS, with frequent and early relapses. NMOSD attacks are often severe resulting in a rapid accumulation of disability (blindness and paraplegia). More effective treatments combined with earlier and more accurate diagnosis has led to improved outcomes. Currently, in the AQP4-IgG era, 5 years after onset, approximately 30% of NMO patients will require a cane to walk and 10% will be wheelchair bound. Treatments for NMOSD include corticosteroids and plasmapheresis for acute attacks and mycophenolate mofetil, azathioprine, and rituximab for relapse prevention. Beta-interferon, a treatment promoted for MS, exacerbates NMOSD. Therefore, early diagnosis and initiation of NMO-appropriate immunosuppressant treatment is important to optimize the clinical outcome by preventing further attacks. Skeletal muscle abnormalities with hyperCKemia have been reported in a few NMOSD patients. Recent reports indicate focal retinal vascular attenuation, inner nuclear layer thickening and microcystic edema in some NMO patients. The sensitivity and specificity of Fluorescence-Activated Cell Sorting (FACS) assay for NMO is >80% and >99%, respectively.

 

Detection of NMO/APQ4-IgG allows distinction of NMOSD from MS and is indicative of a relapsing disease, mandating initiation of immunosuppression, even after the first attack, thereby reducing attack frequency and disability in the future.

Reference Values Describes reference intervals and additional information for interpretation of test results. May include intervals based on age and sex when appropriate. Intervals are Mayo-derived, unless otherwise designated. If an interpretive report is provided, the reference value field will state this.

Negative

Interpretation Provides information to assist in interpretation of the test results

A positive value is consistent with a neuromyelitis optica spectrum disorder (NMOSD) and justifies initiation of appropriate immunosuppressive therapy at the earliest possible time. This allows early initiation and maintenance of optimal therapy. Recommend follow-up in 3 to 6 months if NMOSD is suspected.

 

This autoantibody is not found in healthy subjects.

Cautions Discusses conditions that may cause diagnostic confusion, including improper specimen collection and handling, inappropriate test selection, and interfering substances

AQP4-IgG antibodies may drop below detectable levels in setting of therapies for acute attack (IV methylprednisolone or plasmapheresis) or attack prevention (immunosuppressants).

Supportive Data

An international collaborative group (Mayo Clinic, Oxford University, and McGill University) compared sensitivity and specificity of AQP4 FACS assays to other assay types including fixed, permeabilized cell-based assays (CBA, observer-scored immunofluorescence, Euroimmun), tissue-based immunofluorescence (IF), ELISA, and immunoprecipitation assay (IPA) in a blinded fashion among 60 neuromyelitis optica spectrum disorder (NMOSD) cases and 86 control subjects. Clinical sensitivity of AQP4 FACS was superior to the other assay types. Sensitivities were: AQP4 FACS (M23 isoform), 77%, AQP4-CBA (M1 isoform), 73%; M1-AQP4-ELISA, 60%; IPA, 53%; tissue-based IF, 48%. Specificities were 100% for all assay types, except the Mayo Clinic IPA (97%).(6)

 

In 2014, a systematic comparison of AQP4-IgG assays, in a clinical service setting, confirmed superiority of FACS assays over ELISA. Higher-order arrays of M23-AQP4 (M23-AQP4-FACS) and M1-AQP4-ELISA were associated with false-positive results. Overall, M1-AQP4-FACS was 83% sensitive for NMO compared with 75% for M23-AQP4-FACS, 75% for M1-AQP4-CBA and 58% for M1-AQP4-ELISA. Assays specificities for NMO were: M1-AQP4-FACS, 100%, M1-AQP4-CBA, 100%, M1-AQP4- ELISA, 99%; and M23-AQP4- FACS, 95%.(7)

 

AQP4 FACS analysis was done for serum samples from 36 random patients with a diagnosis of NMO. All samples were tested with our validated AQP4 CBA assay. Thirty samples (83.33%) were positive by FACS and 29 samples (80.55%) were positive by CBA. All 6 samples that were negative by FACS also tested negative by CBA.

 

To measure the specificity, AQP4 FACS analysis was done for CSF of 338 non-NMO(SD) patients. None of the samples were positive by this assay (specificity=100%).

Clinical Reference Recommendations for in-depth reading of a clinical nature

1. Wingerchuk DM, Lennon VA, Lucchinetti CF, et al: The spectrum of neuromyelitis optica. Lancet Neurol 2007;6:805-815

2. Lennon VA, Wingerchuk DM, Kryzer TJ, et al: A serum autoantibody marker of neuromyelitis optica; distinction from multiple sclerosis. Lancet 2004;364:2106-2112

3. Pittock SJ, Weinshenker BG, Lucchinetti CF, et al: Neuromyelitis optica brain lesions localized at sites of high aquaporin 4 expression. Arch Neurol 2006 Jul;63(7):964-968

4. McKeon A, Lennon VA, Lotze T, et al: CNS aquaporin-4 autoimmunity in children. Neurology 2008 Jul 8;71(2):93-100

5. Apiwattanakul M, Popescu BF, Matiello M, et al: Intractable vomiting as the initial presentation of NMO. Ann Neurol 2010 Nov;68(5):757-761

6. Waters P, McKeon A, Leite MI, et al: Multicentre comparison of aquaporin-4 IgG assays in NMO spectrum disorders. Neurology 2012;78:665-671

7. Fryer JP, Lennon VA, Pittock SJ, et al: AQP4 autoantibody assay performance in clinical laboratory service. Neurol Neuroimmunol Neuroinflammation 2014;1:e11