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Glioma 2021 WHO Classification: The Superiority of NGS Over IHC in Routine Diagnostics.

Paulina Śledzińska1,2, Marek Bebyn1,3, Ewelina Szczerba1

  • 1Molecular Oncology and Genetics Department, Innovative Medical Forum, The F. Lukaszczyk Oncology Center, 85-796, Bydgoszcz, Poland.

Molecular Diagnosis & Therapy
|September 2, 2022
PubMed
Summary

Accurate IDH1/2 testing for glioma classification is crucial. Next-generation sequencing (NGS) and quantitative polymerase chain reaction (qPCR) are more reliable than immunohistochemistry (IHC) for determining IDH1/2 status in glioma patients.

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Area of Science:

  • Neuro-oncology
  • Molecular Diagnostics
  • Genetics

Background:

  • Accurate detection of genetic variants is critical for glioma patient management, as per the 2021 World Health Organization (WHO CNS5) classification.
  • Key molecular markers include IDH1/2 mutations, TERT substitutions, chromosomal abnormalities, and MGMT promoter methylation.

Purpose of the Study:

  • To evaluate innovative methods for determining IDH1/2 status in glioma patients.
  • To align diagnostic procedures with the latest WHO CNS5 guidelines.

Main Methods:

  • Simultaneous screening of multiple biomarkers using next-generation sequencing (NGS) on 34 glioma samples.
  • Validation of discrepant results using quantitative polymerase chain reaction (qPCR) and Sanger sequencing when IDH1/2 status from immunohistochemistry (IHC) or multiplex ligation-dependent probe amplification (MLPA) differed from NGS.

Main Results:

  • Significant discrepancies were observed between NGS and IHC (7/13 samples) and NGS and MLPA (1/4 samples) for IDH1/2 status.
  • All NGS findings were confirmed by qPCR and Sanger sequencing, highlighting the reliability of these methods.
  • The WHO CNS5 classification necessitates the assessment of multiple mutations for accurate glioma classification.

Conclusions:

  • Quantitative polymerase chain reaction (qPCR) or NGS performed in reference genetic laboratories are the most reliable methods for IDH1/2 analysis in glioma.
  • Clinicians should be aware of potential discrepancies with IHC or MLPA and seek advanced molecular testing when necessary.
  • A new algorithm for molecular diagnostic procedures in glioma patients, based on WHO CNS5, was proposed.