Contributions
Abstract: P330
Type: Poster Sessions
Abstract Category: Clinical aspects of MS - Diagnosis and differential diagnosis
Background: Antibodies to myelin-oligodendrocyte-glycoprotein (MOG-ab) are associated with a broad spectrum of human CNS demyelinating diseases including AQP4-ab-seronegative neuromyelitis optica spectrum disorders and related clinical presentations such as isolated optic neuritis and myelitis, acute disseminated encephalomyelitis, but only rarely in multiple sclerosis. Although numerous studies have used different immunoassays for MOG-ab, there is no published blinded multi-centre validation experiments comparing MOG-ab assays. The aim of our study was therefore to compare the most frequently used assays for MOG-ab such as immunofluorescence cell-based assays (IF-CBA), flow cytometry cell-based assays (FACS-CBA) and enzyme-linked immunosorbent assay (ELISA). In phase one we compared 11 different MOG-ab assays (in-house and commercial; IF-CBA, FACS-CBA and ELISA) in a blinded fashion on 89 serum samples.
Methods: The clinical laboratories (Innsbruck, Mayo, Oxford and Sydney) sent coded MOG-ab positive sera (n=39), MOG-ab negative sera (n=40) and clinical documentation to the Institute for Quality Assurance (IQA), Lübeck, Germany. Euroimmun AG contributed 10 technical controls (humanized monoclonal antibodies) which were also sent to the IQA. All samples and controls were re-coded, aliquoted and distributed to the five testing centres. Upon completion of the testing, the assay results from each centre was entered onto a web-based database. The data were then unblinded, and analysed.
Results: We found a very good agreement between live IF-CBA and live FACS-CBA (kappa values > 0.9). There was a good agreement between live CBA (IF or FACS) and fixed IF-CBA (kappa values > 0.8). The agreement of ELISA and all CBA was very poor and our results indicate that ELISA is not useful for the detection of human MOG-ab. However, phase I only focused on clearly positive and negative samples, and we are therefore currently performing phase II using 100 additional samples (borderline and lower positive, healthy controls, and replicates from phase I) to compare the sensitivity and specificity of assays. We plan to perform a phase III to examine MOG-ab assays in routine diagnostic centres world-wide and are particularly interested in centres that can contribute sera to the study.
Conclusions: Large multicentre validation studies of antibody assay are important to evaluate assay reproducibility between centres and to help define the clinical phenotype associated with MOG-ab.
Disclosure: Markus Reindl was supported by a research grant from the Austrian Science Promotion Agency (FFG). The University Hospital and Medical University of Innsbruck (Austria; M.R.) receives payments for antibody assays (MOG, AQP4, and other autoantibodies) and for MOG and AQP4 antibody validation experiments organized by Euroimmun (Lübeck, Germany).
Kathrin Schanda was supported by a research grant from the Austrian Science Promotion Agency (FFG).
Mark Woodhall has no disclosure to report.
Fiona Tea has no disclosure to report.
Sudarshini Ramanathan has no disclosure to report.
Bianca Teegen has received personal compensation from Labor Dr. Stoecker as an employee.
Jessica Sagen has no disclosure to report.
Jim Fryer has no disclosure to report.
John Mills has no disclosure to report.
Swantje Mindorf has received personal compensation from Euroimmun AG as an employee.
Nora Ritter has received personal compensation from Euroimmun AG as an employee.
Ulrike Krummrei has received personal compensation from Euroimmun AG as an employee.
Winfried Stöcker has received personal compensation from Euroimmun AG as CEO.
Juliane Eggert has received personal compensation from Euroimmunas an employee of the Institute for Quality Assurance.
Kevin Rostasy has no disclosure to report.
Thomas Berger: the University Hospital and Medical University of Innsbruck (Austria; T.B.) receives payments for antibody assays (MOG, AQP4, and other autoantibodies) and for MOG and AQP4 antibody validation experiments organized by Euroimmun (Lübeck, Germany).
Sarosh R Irani reports personal fees from MedImmune and has a patent WO/2010/046716 entitled 'Neurological Autoimmune Disorders' with royalties paid.
Russell Dale was supported by research grants from the National Health and Medical Research Council (NHRMC), Multiple Sclerosis Research Australia (MSRA), and Trish Multiple Sclerosis Research Foundation.
Christian Probst has received personal compensation from Euroimmun AG as an employee.
Fabienne Brilot was supported by research grants from the National Health and Medical Research Council (NHRMC), Multiple Sclerosis Research Australia (MSRA), and Trish Multiple Sclerosis Research Foundation.
Monika Probst has received personal compensation from Euroimmunas an employee of the Institute for Quality Assurance.
Sean Pittock is a named inventor on filed patents that relate to functional AQP4/NMO-IgG assays and NMO-IgG as a cancer marker; consulted for Alexion, Medimmune, Grifols and Euroimmun; and received research support from Grifols, Medimmune, Alexion, Euroimmun, and AEA (Autoimmune Encephalitis Alliance). All compensation for consulting activities is paid directly to Mayo Clinic.
Patrick Waters and the University of Oxford are named inventors on patents for antibody assays and have received royalties. He has received honoraria or research funding from Biogen Idec, Mereo biopharma, Retrogenix and Euroimmun AG, and travel grants from the Guthy-Jackson Charitable Foundation.
Abstract: P330
Type: Poster Sessions
Abstract Category: Clinical aspects of MS - Diagnosis and differential diagnosis
Background: Antibodies to myelin-oligodendrocyte-glycoprotein (MOG-ab) are associated with a broad spectrum of human CNS demyelinating diseases including AQP4-ab-seronegative neuromyelitis optica spectrum disorders and related clinical presentations such as isolated optic neuritis and myelitis, acute disseminated encephalomyelitis, but only rarely in multiple sclerosis. Although numerous studies have used different immunoassays for MOG-ab, there is no published blinded multi-centre validation experiments comparing MOG-ab assays. The aim of our study was therefore to compare the most frequently used assays for MOG-ab such as immunofluorescence cell-based assays (IF-CBA), flow cytometry cell-based assays (FACS-CBA) and enzyme-linked immunosorbent assay (ELISA). In phase one we compared 11 different MOG-ab assays (in-house and commercial; IF-CBA, FACS-CBA and ELISA) in a blinded fashion on 89 serum samples.
Methods: The clinical laboratories (Innsbruck, Mayo, Oxford and Sydney) sent coded MOG-ab positive sera (n=39), MOG-ab negative sera (n=40) and clinical documentation to the Institute for Quality Assurance (IQA), Lübeck, Germany. Euroimmun AG contributed 10 technical controls (humanized monoclonal antibodies) which were also sent to the IQA. All samples and controls were re-coded, aliquoted and distributed to the five testing centres. Upon completion of the testing, the assay results from each centre was entered onto a web-based database. The data were then unblinded, and analysed.
Results: We found a very good agreement between live IF-CBA and live FACS-CBA (kappa values > 0.9). There was a good agreement between live CBA (IF or FACS) and fixed IF-CBA (kappa values > 0.8). The agreement of ELISA and all CBA was very poor and our results indicate that ELISA is not useful for the detection of human MOG-ab. However, phase I only focused on clearly positive and negative samples, and we are therefore currently performing phase II using 100 additional samples (borderline and lower positive, healthy controls, and replicates from phase I) to compare the sensitivity and specificity of assays. We plan to perform a phase III to examine MOG-ab assays in routine diagnostic centres world-wide and are particularly interested in centres that can contribute sera to the study.
Conclusions: Large multicentre validation studies of antibody assay are important to evaluate assay reproducibility between centres and to help define the clinical phenotype associated with MOG-ab.
Disclosure: Markus Reindl was supported by a research grant from the Austrian Science Promotion Agency (FFG). The University Hospital and Medical University of Innsbruck (Austria; M.R.) receives payments for antibody assays (MOG, AQP4, and other autoantibodies) and for MOG and AQP4 antibody validation experiments organized by Euroimmun (Lübeck, Germany).
Kathrin Schanda was supported by a research grant from the Austrian Science Promotion Agency (FFG).
Mark Woodhall has no disclosure to report.
Fiona Tea has no disclosure to report.
Sudarshini Ramanathan has no disclosure to report.
Bianca Teegen has received personal compensation from Labor Dr. Stoecker as an employee.
Jessica Sagen has no disclosure to report.
Jim Fryer has no disclosure to report.
John Mills has no disclosure to report.
Swantje Mindorf has received personal compensation from Euroimmun AG as an employee.
Nora Ritter has received personal compensation from Euroimmun AG as an employee.
Ulrike Krummrei has received personal compensation from Euroimmun AG as an employee.
Winfried Stöcker has received personal compensation from Euroimmun AG as CEO.
Juliane Eggert has received personal compensation from Euroimmunas an employee of the Institute for Quality Assurance.
Kevin Rostasy has no disclosure to report.
Thomas Berger: the University Hospital and Medical University of Innsbruck (Austria; T.B.) receives payments for antibody assays (MOG, AQP4, and other autoantibodies) and for MOG and AQP4 antibody validation experiments organized by Euroimmun (Lübeck, Germany).
Sarosh R Irani reports personal fees from MedImmune and has a patent WO/2010/046716 entitled 'Neurological Autoimmune Disorders' with royalties paid.
Russell Dale was supported by research grants from the National Health and Medical Research Council (NHRMC), Multiple Sclerosis Research Australia (MSRA), and Trish Multiple Sclerosis Research Foundation.
Christian Probst has received personal compensation from Euroimmun AG as an employee.
Fabienne Brilot was supported by research grants from the National Health and Medical Research Council (NHRMC), Multiple Sclerosis Research Australia (MSRA), and Trish Multiple Sclerosis Research Foundation.
Monika Probst has received personal compensation from Euroimmunas an employee of the Institute for Quality Assurance.
Sean Pittock is a named inventor on filed patents that relate to functional AQP4/NMO-IgG assays and NMO-IgG as a cancer marker; consulted for Alexion, Medimmune, Grifols and Euroimmun; and received research support from Grifols, Medimmune, Alexion, Euroimmun, and AEA (Autoimmune Encephalitis Alliance). All compensation for consulting activities is paid directly to Mayo Clinic.
Patrick Waters and the University of Oxford are named inventors on patents for antibody assays and have received royalties. He has received honoraria or research funding from Biogen Idec, Mereo biopharma, Retrogenix and Euroimmun AG, and travel grants from the Guthy-Jackson Charitable Foundation.