Diagnostic value of anti-GM1 ganglioside serology and validation of the INCAT-ELISA

doi:10.1016/j.jns.2005.07.009

Journal of the Neurological Sciences

Volume 239, Issue 1, 15 December 2005, Pages 37-44

https://doi.org/10.1016/j.jns.2005.07.009 Get rights and content

Abstract

The Inflammatory Neuropathy and Treatment (INCAT) group developed a standardized ELISA method for the detection of serum anti-GM1 antibodies. The diagnostic value of anti-GM1 antibodies determined by this method has not yet been established in large groups of patients. We assessed the reproducibility, sources of variation, optimal cut-off values and evaluated the diagnostic relevance of the INCAT-ELISA in various groups of patients and controls (N = 1232). The coefficient of variance was 11.2% for IgM and 3.8% for IgG. High IgG titers were only found in Guillain–Barré syndrome (GBS) and other inflammatory polyneuropathies. High IgM titers were associated with GBS and multifocal motor neuropathy. Low IgM titers had no additional diagnostic value. The INCAT-ELISA is a reliable test with additional diagnostic value in specific clinical situations.

Introduction

Determining antibodies to the ganglioside GM1 as a diagnostic marker for immune mediated neuropathies remains controversial because they have been reported in a wide spectrum of neurological diseases, autoimmune disorders and in healthy controls. The frequency of anti-GM1 antibodies in these groups of patients and controls also varies considerably between reports. These variations could largely be explained by differences in techniques used to detect anti-GM1 antibodies [1].

To facilitate the comparison of results from different laboratories a standardized enzyme-linked immunosorbent assay (ELISA) for measuring serum anti-GM1 antibodies was proposed by the Inflammatory Neuropathy and Treatment (INCAT) group [2]. The INCAT initiative consists of a group of European neurological centers with the aim to develop standardized laboratory and clinical protocols to optimize research, diagnosis and treatment of inflammatory neuropathies. In a previous study of the INCAT-ELISA protocol, the variation between six laboratories was determined. This study showed that 41% of this variation was generated by the intra-laboratory variation [2]. The sources of variation and the frequency of anti-GM1 antibodies in large groups of patients and controls using this standardized INCAT method have not yet been established.

In this study, as an intra-laboratory quality control, the reproducibility and sources of variation of the INCAT-ELISA were established. In addition, the frequency of anti-GM1 antibodies in large groups of patients was determined for the first time using this method. Based on the INCAT-ELISA, this study also provides the positive and negative predictive values for neurological disorders related with anti-GM1 antibodies assessing the diagnostic value of these antibodies in clinical practice.

Section snippets

Patients

Serum samples were obtained from 1232 patients and controls. These serum samples were retrospectively studied for the presence of IgM and IgG anti-GM1 antibodies. The study included patients with multifocal motor neuropathy (MMN, N = 52), Guillain–Barré syndrome (GBS, N = 471), chronic inflammatory demyelinating polyneuropathy (CIDP, N = 82), paraprotein-related polyneuropathy (PP-PNP, N = 102), other inflammatory polyneuropathy (I-PNP, N = 40), non-inflammatory polyneuropathy (NI-PNP, N = 74), motor

Analysis of variance factors in INCAT anti-GM1 ELISA

The results of the experiments to determine the relative contribution of the variation between technicians, days, plates and wells within the plates to the overall variation are shown in Fig. 1. The CV of the overall intra-laboratory assay variance was 11.1% for IgM and 3.8% for IgG. The largest contributive variance factor for IgM was the variation between plates (62% of total variance) and wells within the plates (38% of total variance). For IgG, the largest contributive factor was the

Discussion

The INCAT-ELISA demonstrated good intra-laboratory reproducibility. The overall variability in our laboratory expressed as CV was 11.1% for IgM and 3.8% for IgG, which is lower than the findings in the previous INCAT study [2]. In that study, the intra-laboratory variance was determined less extensively and estimated based on results obtained from various laboratories. In a single experiment, we determined the relative contribution that may influence the intra-laboratory variance of

Acknowledgments

We would like to thank Dr. R Van den Berg-Vos (University Medical Center Utrecht, Utrecht, The Netherlands) for providing serum samples of MMN patients and Dr. M. Eurelings (University Medical Center Utrecht, Utrecht, The Netherlands) for providing serum samples of paraprotein-related polyneuropathy patients. The Erasmus MC, as part of a postgraduate training, financially supported this work.

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Citation Excerpt :
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Diagnostic value of anti-GM1 ganglioside serology and validation of the INCAT-ELISA

Abstract

Introduction

Section snippets

Patients

Analysis of variance factors in INCAT anti-GM1 ELISA

Discussion

Acknowledgments

Lancet

J. Neurol. Sci.

J Neuroimmunol

J Neuroimmunol

J Neuroimmunol

J Neuroimmunol

Autoimmun Rev

Diagnostic value of GM1 antibodies in motor neuron disorders and neuropathies: a meta-analysis

Neurology

Inter-laboratory validation of an ELISA for the determination of serum anti-ganglioside antibodies

Eur J Neurol

A randomized trial comparing intravenous immune globulin and plasma exchange in Guillain–Barré syndrome. Dutch Guillain–Barré Study Group

N Engl J Med