Validation of a Flow Cytometry Live Cell-Based Assay to Detect Myelin Oligodendrocyte Glycoprotein Antibodies for Clinical Diagnostics

Joseph A Lopez^{1

2}, Samuel D Houston^{1

3}, Fiona Tea^{1

2}, Vera Merheb¹, Fiona X Z Lee¹, Sandy Smith⁴, David McDonald⁴, Alicia Zou^{1

2}, Ganesha Liyanage^{1

5}, Deepti Pilli^{1

2}, Martina Denkova^{1

5}, Jeannette Lechner-Scott⁶, Anneke van der Walt⁷, Michael H Barnett⁸, Stephen W Reddel^{8

9}, Simon Broadley^{10

11}, Sudarshini Ramanathan^{1

9

12}, Russell C Dale^{1

2

8}, David A Brown^{4

12

13}, Fabienne Brilot^{1

2

5

8}

Affiliations

¹ Brain Autoimmunity Group, Kids Neuroscience Centre, Kids Research at the Children's Hospital at Westmead, Sydney, Australia.
² Specialty of Child and Adolescent Health, Faculty of Medicine and Health, The University of Sydney, Sydney, Australia.
³ School of Biomedical Engineering, The University of Sydney, Sydney, Australia.
⁴ New South Wales Health Pathology, Institute of Clinical Pathology and Medical Research, Westmead Hospital, Sydney, Australia.
⁵ School of Medical Sciences, Faculty of Medicine and Health, The University of Sydney, Sydney, Australia.
⁶ Hunter Medical Research Institute, Faculty of Medicine and Public Health, The University of Newcastle, Department of Neurology, John Hunter Hospital, Newcastle, Australia.
⁷ Department of Neurosciences, Central Clinical School, Monash University, Melbourne, Australia.
⁸ Brain and Mind Centre, The University of Sydney, Sydney, Australia.
⁹ Department of Neurology, Concord Repatriation General Hospital, Sydney, Australia.
¹⁰ Menzies Health Institute Queensland, Gold Coast Campus, Griffith University Southport, Australia.
¹¹ Department of Neurology, Gold Coast University Hospital, Southport, Australia.
¹² Sydney Medical School, Faculty of Medicine and Health, The University of Sydney, Sydney, Australia.
¹³ Westmead Institute for Medical Research, Sydney, Australia.

PMID: 34718586
DOI: 10.1093/jalm/jfab101

Validation of a Flow Cytometry Live Cell-Based Assay to Detect Myelin Oligodendrocyte Glycoprotein Antibodies for Clinical Diagnostics

Joseph A Lopez et al. J Appl Lab Med. 2022.

. 2022 Jan 5;7(1):12-25.

doi: 10.1093/jalm/jfab101.

Authors

Affiliations

¹ Brain Autoimmunity Group, Kids Neuroscience Centre, Kids Research at the Children's Hospital at Westmead, Sydney, Australia.
² Specialty of Child and Adolescent Health, Faculty of Medicine and Health, The University of Sydney, Sydney, Australia.
³ School of Biomedical Engineering, The University of Sydney, Sydney, Australia.
⁴ New South Wales Health Pathology, Institute of Clinical Pathology and Medical Research, Westmead Hospital, Sydney, Australia.
⁵ School of Medical Sciences, Faculty of Medicine and Health, The University of Sydney, Sydney, Australia.
⁶ Hunter Medical Research Institute, Faculty of Medicine and Public Health, The University of Newcastle, Department of Neurology, John Hunter Hospital, Newcastle, Australia.
⁷ Department of Neurosciences, Central Clinical School, Monash University, Melbourne, Australia.
⁸ Brain and Mind Centre, The University of Sydney, Sydney, Australia.
⁹ Department of Neurology, Concord Repatriation General Hospital, Sydney, Australia.
¹⁰ Menzies Health Institute Queensland, Gold Coast Campus, Griffith University Southport, Australia.
¹¹ Department of Neurology, Gold Coast University Hospital, Southport, Australia.
¹² Sydney Medical School, Faculty of Medicine and Health, The University of Sydney, Sydney, Australia.
¹³ Westmead Institute for Medical Research, Sydney, Australia.

PMID: 34718586
DOI: 10.1093/jalm/jfab101

Abstract

Background: Myelin oligodendrocyte glycoprotein antibodies (MOG Ab) are essential in the diagnosis of MOG Ab-associated disease (MOGAD). Live cell-based assays (CBAs) are the gold standard for MOG Ab detection with improved sensitivity and specificity over fixed CBAs. A number of testing centers have used flow cytometry for its high throughput and quantitative utility. Presently, there is increasing demand to translate these research-based methods into an accredited routine diagnostic setting.

Methods: A flow cytometry live CBA was used to detect MOG Ab in patients with demyelination. Serostatuses were compared between a research-based assay and a streamlined diagnostic assay. Inter-laboratory validation of the streamlined assay was performed in an accredited diagnostic laboratory. Further streamlining was performed by introducing a borderline serostatus range and reducing the number of controls used to determine the positivity threshold.

Results: High serostatus agreement (98%-100%) was observed between streamlined and research-based assays. Intra- and inter-assay imprecision was improved in the streamlined assay (mean intra- and inter-assay CV = 7.3% and 27.8%, respectively) compared to the research-based assay (mean intra- and inter-assay CV = 11.8% and 33.6%, respectively). Borderline positive and clear positive serostatuses were associated with confirmed phenotypes typical of MOGAD. Compared to using 24 controls, robust serostatus classification was observed when using 13 controls without compromising analytical performance (93%-98.5% agreement).

Conclusions: Flow cytometry live CBAs show robust utility in determining MOG Ab serostatus. Streamlining and standardizing use of this assay for diagnostics would improve the accuracy and reliability of routine testing to aid diagnosis and treatment of patients with demyelination.

Keywords: MOG antibody; demyelinating disorders; diagnostic validation; flow cytometry.

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Validation of a Flow Cytometry Live Cell-Based Assay to Detect Myelin Oligodendrocyte Glycoprotein Antibodies for Clinical Diagnostics

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Validation of a Flow Cytometry Live Cell-Based Assay to Detect Myelin Oligodendrocyte Glycoprotein Antibodies for Clinical Diagnostics

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Abstract

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