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Multiplexing analysis of the polyspecific intrathecal immune response in multiple sclerosis
Alina Kułakowska1, Barbara Mroczko, Maria Mantur
1Department of Neurology, Medical University of Białystok, Poland.
Abstract:
Intrathecal synthesis of the antibodies specific to neurotrofic viruses: measles (M), rubella (R), Varicella-Zoster (Z), and/or H. simplex (H), known as "MRZH-reaction" plays important diagnostic role in multiple sclerosis (MS). Whereas the analysis of the oligoclonal IgG bands provides high sensitivity, the MRZH-reaction shows high specificity, and hence these methods complement each other. For the first time we applied multiplexing bead-based technology to simultaneously analyze cerebrospinal fluid (CSF) and serum concentrations of antibodies against these viruses, and to calculate the antibody specific indices (ASI's). The method shows reasonable precision: intra-assay, 2.9-6.7%, and inter-assay, 2.0-3.2%. The results are comparable with these obtained with other methods (ELISAs), including two runs of the certified external quality control schemes. Eighty-one percent of the MS cases (n=27) and none of the sex- and age-matched controls (n=14), except one subject with "borderline" anti-measles ASI of 1.5, showed intrathecal synthesis of IgG against at least one of the viruses discussed. The ratios of the MRZH-positive cases in the MS group were: 12/22 for M, 12/19 for R, 13/26 for Z, and 7/26 for H. We conclude that the multiplexing technology can be applied as a tool to study the intrathecal immune response in the diagnosis of MS.
Insights
Multiplexing technology accurately detects intrathecal antibody synthesis against measles, rubella, Varicella-Zoster, and herpes simplex viruses in multiple sclerosis (MS) patients. This method enhances diagnostic specificity for MS by identifying the MRZH-reaction.
Area of Science:
- Neuroimmunology
- Virology
- Diagnostic Technology
Background:
- The "MRZH-reaction" (measles, rubella, Varicella-Zoster, herpes simplex virus antibodies) is crucial for diagnosing multiple sclerosis (MS).
- While oligoclonal IgG bands offer high sensitivity, the MRZH-reaction provides high specificity, making them complementary diagnostic tools.
Purpose of the Study:
- To evaluate multiplexing bead-based technology for simultaneous analysis of cerebrospinal fluid (CSF) and serum antibodies against neurotrophic viruses (measles, rubella, Varicella-Zoster, herpes simplex).
- To assess the utility of this technology in calculating antibody specific indices (ASIs) for diagnosing MS.
Main Methods:
- Application of multiplexing bead-based technology to quantify antibodies against measles, rubella, Varicella-Zoster, and herpes simplex viruses in CSF and serum.
- Calculation of antibody specific indices (ASIs) to determine intrathecal antibody synthesis.
- Validation of the method's precision (intra-assay 2.9-6.7%, inter-assay 2.0-3.2%) and comparison with ELISA.
Main Results:
- Multiplexing technology demonstrated good precision and comparable results to ELISAs.
- Eighty-one percent of MS patients (n=27) exhibited intrathecal IgG synthesis against at least one of the studied viruses.
- No healthy controls (n=14) showed significant intrathecal synthesis, except one borderline case.
Conclusions:
- Multiplexing bead-based technology is a precise and effective tool for studying intrathecal immune responses in MS.
- This technology aids in diagnosing MS by identifying the MRZH-reaction, complementing existing methods like oligoclonal band analysis.
