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Related Concept Videos

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Optimization, Design and Avoiding Pitfalls in Manual Multiplex Fluorescent Immunohistochemistry
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An optimized immunohistochemistry technique improves NMO-IgG detection: study comparison with cell-based assays.

Romana Höftberger1, Lidia Sabater, Romain Marignier

  • 1Service of Neurology, Hospital Clínic, Universitat de Barcelona and Institut d´Investigació Biomèdica August Pi i Sunyer (IDIBAPS), Barcelona, Spain ; Institute of Neurology, Medical University of Vienna, Vienna, Austria.

Plos One
|November 14, 2013
PubMed
Summary

An optimized immunohistochemistry assay shows comparable sensitivity and specificity to cell-based assays for detecting neuromyelitis optica (NMO)-IgG/aquaporin-4-antibody. This finding offers a valuable alternative tool for NMO diagnosis.

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Area of Science:

  • Neurology
  • Immunology
  • Diagnostic Assays

Background:

  • Cell-based assays (CBA) have improved detection of neuromyelitis optica (NMO)-IgG/aquaporin-4-antibody.
  • Classical tissue-based assays have limitations in sensitivity.

Purpose of the Study:

  • To compare the sensitivity and specificity of an optimized immunohistochemistry (IHC-o) assay.
  • To evaluate IHC-o against in-house (CBA-ih) and commercial (CBA-c) cell-based assays for NMO-IgG/aquaporin-4-antibody detection.

Main Methods:

  • Evaluated coded serum from 103 patients with definite NMO and 122 controls.
  • Utilized IHC-o with a protocol for cell surface antigens.
  • Employed CBA-ih with live cells expressing aquaporin-4-M23-isoform and CBA-c.

Main Results:

  • IHC-o sensitivity was 74.8%, comparable to CBA-ih (75.7%) and CBA-c (73.8%).
  • All three assays demonstrated 100% specificity.
  • High interassay concordance was observed, with 100 of 103 samples showing agreement.

Conclusions:

  • Optimized immunohistochemistry is a sensitive and specific method for NMO-IgG/aquaporin-4-antibody detection.
  • IHC-o serves as a valuable addition to existing diagnostic tools for NMO.