Paulina Śledzińska1,2, Marek Bebyn1,3, Ewelina Szczerba1, Jacek Furtak4,5, Maciej Harat5,6, Natalia Olszewska1, Katarzyna Kamińska1, Janusz Kowalewski2, Marzena A Lewandowska7,8. 1. Molecular Oncology and Genetics Department, Innovative Medical Forum, The F. Lukaszczyk Oncology Center, 85-796, Bydgoszcz, Poland. 2. Department of Thoracic Surgery and Tumors, Ludwik Rydygier Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University, 85-067, Torun, Poland. 3. Faculty of Medicine, Medical University of Gdańsk, 80-210, Gdańsk, Poland. 4. Department of Neurosurgery, 10th Military Research Hospital and Polyclinic, 85-681, Bydgoszcz, Poland. 5. Department of Neurooncology and Radiosurgery, The F. Lukaszczyk Oncology Center, 85-796, Bydgoszcz, Poland. 6. Department of Oncology and Brachytherapy, Ludwik Rydygier Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University, 85-067, Bydgoszcz, Poland. 7. Molecular Oncology and Genetics Department, Innovative Medical Forum, The F. Lukaszczyk Oncology Center, 85-796, Bydgoszcz, Poland. lewandowskam@co.bydgoszcz.pl. 8. Department of Thoracic Surgery and Tumors, Ludwik Rydygier Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University, 85-067, Torun, Poland. lewandowskam@co.bydgoszcz.pl.
Abstract
INTRODUCTION: The accurate detection of genetic variants such as single substitutions (IDH1/2, TERT), chromosomal abnormalities (CDKN2A, 1p/19q deletions, and EGFR amplifications), or promoter methylations (MGMT) is critical for glioma patient management, as emphasized in the World Health Organization's (WHO's) most recent classification in 2021 (WHO CNS5). The purpose of this study was to evaluate novel innovative methods for determining IDH1/2 status in the context of WHO CNS5. METHODS: Multiple biomarkers were simultaneously screened using next-generation sequencing (NGS) on 34 glioma samples. In cases where the IDH1/2 status determined by immunohistochemistry (IHC) or multiplex ligation-dependent probe amplification (MLPA) was inconsistent with the NGS results, quantitative polymerase chain reaction (qPCR) and Sanger sequencing were performed to resolve the adjudicated discrepancy. RESULTS: IDH1/2 NGS results differ from IHC (7/13 samples) as well as MLPA reports (1/4 samples). All NGS findings were confirmed by qPCR and Sanger sequencing. WHO CNS5 requires assessment of multiple mutations for glioma classification. CONCLUSIONS: We demonstrated that qPCR or NGS performed in reference genetic laboratories, rather than IHC, is the most reliable method for IDH1/2 analysis. Clinicians should be aware of discrepancies in MLPA or IHC results and seek reconsultation in facilities with extensive access to advanced molecular technologies. Moreover, we proposed a new algorithm for the molecular diagnostic procedures in glioma patients based on the WHO CNS5.
INTRODUCTION: The accurate detection of genetic variants such as single substitutions (IDH1/2, TERT), chromosomal abnormalities (CDKN2A, 1p/19q deletions, and EGFR amplifications), or promoter methylations (MGMT) is critical for glioma patient management, as emphasized in the World Health Organization's (WHO's) most recent classification in 2021 (WHO CNS5). The purpose of this study was to evaluate novel innovative methods for determining IDH1/2 status in the context of WHO CNS5. METHODS: Multiple biomarkers were simultaneously screened using next-generation sequencing (NGS) on 34 glioma samples. In cases where the IDH1/2 status determined by immunohistochemistry (IHC) or multiplex ligation-dependent probe amplification (MLPA) was inconsistent with the NGS results, quantitative polymerase chain reaction (qPCR) and Sanger sequencing were performed to resolve the adjudicated discrepancy. RESULTS: IDH1/2 NGS results differ from IHC (7/13 samples) as well as MLPA reports (1/4 samples). All NGS findings were confirmed by qPCR and Sanger sequencing. WHO CNS5 requires assessment of multiple mutations for glioma classification. CONCLUSIONS: We demonstrated that qPCR or NGS performed in reference genetic laboratories, rather than IHC, is the most reliable method for IDH1/2 analysis. Clinicians should be aware of discrepancies in MLPA or IHC results and seek reconsultation in facilities with extensive access to advanced molecular technologies. Moreover, we proposed a new algorithm for the molecular diagnostic procedures in glioma patients based on the WHO CNS5.
Authors: Athanasios C Tsiatis; Alexis Norris-Kirby; Roy G Rich; Michael J Hafez; Christopher D Gocke; James R Eshleman; Kathleen M Murphy Journal: J Mol Diagn Date: 2010-04-29 Impact factor: 5.568
Authors: Ewelina Szczerba; Katarzyna Kamińska; Tomasz Mierzwa; Marcin Misiek; Janusz Kowalewski; Marzena Anna Lewandowska Journal: Genes (Basel) Date: 2021-04-02 Impact factor: 4.096
Authors: Marzena Anna Lewandowska; Jacek Furtak; Tadeusz Szylberg; Krzysztof Roszkowski; Wiesława Windorbska; Joanna Rytlewska; Wojciech Jóźwicki Journal: Mol Diagn Ther Date: 2014-02 Impact factor: 4.074