Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
Comparative Study
. 2025 Dec 15:409:578760.
doi: 10.1016/j.jneuroim.2025.578760. Epub 2025 Sep 17.

Live MOG-IgG cell-based assay: Comparison across flow cytometers and diagnostic validation on high-sensitivity full spectrum flow cytometry

Elisha Siwan  1 , Asawin Tungpoomjaruswong  2 , Vera Merheb  3 , Fiona X Z Lee  3 , Fakhria Kakar  4 , Mark Acebes  2 , Russell C Dale  1 , David McDonald  2 , Sudarshini Ramanathan  5 , Ming-Wei Lin  6 , David A Brown  6 , Fabienne Brilot  7
Affiliations
Free article
Comparative Study

Live MOG-IgG cell-based assay: Comparison across flow cytometers and diagnostic validation on high-sensitivity full spectrum flow cytometry

Elisha Siwan et al. J Neuroimmunol. .
Free article

Abstract

MOG antibody-associated disease (MOGAD) diagnosis rests on seropositivity for MOG antibody (MOG-IgG). Live cell-based assays (CBA) are gold standards. Although flow cytometry live CBAs have high real-world sensitivity, their global implementation and diagnostic deployment have been challenged by perceptions of "in-house" design and custom optimization. Herein, we compared the analytical robustness of flow live MOG-IgG CBA across various cytometers in both research and diagnostic laboratories. Flow live CBAs were performed on three conventional (Fortessa, BDLSRII, Gallios), and two spectral cytometers (Aurora, ID7000). MOG-IgG titers were calculated by median fluorescence intensity (MFI), and intra- and inter assay precisions (CV%) and serostatuses were determined. The MFI detection range on Fortessa, currently used for testing, was significantly lower than spectral ID7000 (4.75-fold, p = 0.04) and Aurora (12-fold, p = 0.0001), albeit all MFIs correlated (p < 0.0001; R2 = 0.99). Interestingly, the high detection range was attributed to technology, not laboratory environments (p > 0.05). Intra- and inter-assay precisions were similar across cytometers. ID7000 and Fortessa had the lowest variation, with 4.6 % and 6.8 % intra-CV, respectively, and 15.7 % inter-CV. FACS ratio, currently reported as MOG-IgG titre, and MFIs were comparable and correlated for all cytometers (p < 0.001; R2 ≤ 0.99), regardless of the analysis software (p < 0.0001, R2 = 0.98). All serostatuses were highly concordant (κ = 1). Our results demonstrate that flow live CBAs can be validated in diagnostic laboratories across a range of flow cytometers with high reproducibility and repeatability. In particular, excellent assay performance on spectral flow cytometry strongly supports the proof-of-concept use of this technology for diagnostic purposes.

Keywords: Demyelinating disorders; Diagnostic validation; Flow cytometry; Live cell-based assay; MOG antibody; MOGAD; Validation.

PubMed Disclaimer

Conflict of interest statement

Declaration of competing interest ES, AT, VM, FXZL, FK, MA, DmcD, and MWL have no conflict of interest. RCD has received research funding from the Star Scientific Foundation, The Trish Multiple Sclerosis Research Foundation, Multiple Sclerosis Research Australia, the Petre Foundation and the NHMRC (Australia; Investigator Grant). He has also received honoraria from Biogen Idec as an invited speaker, and is on the IDMC for a Roche RCT in paediatric MS. He is on the medical advisory board (non-remunerated position). SR received research funding from the National Health and Medical Research Council (Australia), the Royal Australasian College of Physicians, and the University of Sydney. She serves as a consultant on an advisory board for UCB and Limbic Neurology, The MOG Project and the Sumaira Foundation, and has been an invited speaker for Biogen, Excemed, Alexxion, Limbic Neurology, and Novartis. DAB receives royalties from Roche Diagnostics for assays commercialised by St Vincent's hospital, Sydney, Australia. FB has received research funding from investigator-initiated research grant from Novartis and MS Australia for unrelated projects. She also received funding from NSW Health, the National Health Medical Research Council (Australia), the Medical Research Future Fund (Australia). She was on advisory boards for Novartis, Merck, and The MOG Project and the Sumaira Foundation, and has been an invited speaker for UCB, Biogen, Novartis, and Limbic Neurology. She leads the partnership between the University of Sydney, the Children's Hospital at Westmead, and New South Wales Health Pathology to offer accredited MOG-IgG testing (Westmead, Sydney, New South Wales, Australia).

LinkOut - more resources