Standardized method for the detection of antibodies to aquaporin-4 based on a highly sensitive immunofluorescence assay employing recombinant target antigen

Background - Recently, a highly specific serum autoantibody was discovered in patients with neuromyelitis optica, called NMO-IgG, and aquaporin-4, the most abundant water channel in the CNS, was identified as the target antigen. Several assays for the detection of NMO-IgG/AQP4-Ab have been described...

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Hauptverfasser: Jarius, Sven (VerfasserIn) , Probst, C. (VerfasserIn) , Borowski, K. (VerfasserIn) , Franciotta, D. (VerfasserIn) , Wildemann, Brigitte (VerfasserIn) , Stoecker, W. (VerfasserIn) , Wandinger, K. P. (VerfasserIn)
Dokumenttyp: Article (Journal)
Sprache:Englisch
Veröffentlicht: [15 April 2010]
In: Journal of the neurological sciences
Year: 2010, Jahrgang: 291, Heft: 1, Pages: 52-56
ISSN:1878-5883
DOI:10.1016/j.jns.2010.01.002
Online-Zugang:Verlag, lizenzpflichtig, Volltext: https://doi.org/10.1016/j.jns.2010.01.002
Verlag, lizenzpflichtig, Volltext: https://www.sciencedirect.com/science/article/pii/S0022510X10000109
Volltext
Verfasserangaben:S. Jarius, C. Probst, K. Borowski, D. Franciotta, B. Wildemann, W. Stoecker, K.P. Wandinger

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520 |a Background - Recently, a highly specific serum autoantibody was discovered in patients with neuromyelitis optica, called NMO-IgG, and aquaporin-4, the most abundant water channel in the CNS, was identified as the target antigen. Several assays for the detection of NMO-IgG/AQP4-Ab have been described. Tests based on recombinant human AQP4 have been repeatedly demonstrated to be more sensitive than the previous gold standard assay, i.e. immunohistochemistry (IHC) on mouse brain tissue. However, the sophisticated techniques applied restrict their availability to few laboratories worldwide. - Objective - To develop an easy-to-use, recombinant immunofluorescence assay (rIFA) suitable for standardized and high-throughput detection of NMO-IgG/AQP4-Ab. - Methods - HEK293 cells seeded on cover glasses were transfected with full-length recombinant human AQP4 at large scale. Cover glasses with the immobilized cells were cut into millimetre-sized fragments and transferred to microscopy slides. 151 serum samples from patients with NMO spectrum disorders (NMOSD) and controls were analysed both in the standard IHC assay and in the newly developed rIFA. - Results - 25/32 (78.1%) patients with clinically definite NMO and 36/51 (70.6%) of total patients with NMOSD were positive for NMO-IgG/AQP4-Ab in the rIFA compared to 65.6% and 58.8%, respectively, in the IHC assay. - Conclusion - The recombinant IFA presented here provides laboratories familiar with indirect immunofluorescence microscopy with a highly sensitive and reproducible diagnostic tool for standardized detection of antibodies to AQP4. This new approach could make AQP4-Ab testing, which is of high clinical relevance, more widely available. 
650 4 |a Antibody to aquaporin-4 
650 4 |a Diagnosis 
650 4 |a Immunofluorescence assay 
650 4 |a Longitudinally extensive transverse myelitis 
650 4 |a Multiple sclerosis 
650 4 |a Neuromyelitis optica (Devic syndrome) 
650 4 |a NMO-IgG 
650 4 |a Optic neuritis 
650 4 |a Recombinant antigen 
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700 1 |a Stoecker, W.  |e VerfasserIn  |4 aut 
700 1 |a Wandinger, K. P.  |e VerfasserIn  |4 aut 
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