Multiplexed Droplet Digital PCR Assays for the Simultaneous Screening of Major Genetic Alterations in Tumors of the Central Nervous System.
biomarkers
formaldehyde-fixed sample tissue
glial and glioneuronal tumors
molecular screening test
multiplexed droplet digital PCR assay
tumors of the central nervous system
Journal
Frontiers in oncology
ISSN: 2234-943X
Titre abrégé: Front Oncol
Pays: Switzerland
ID NLM: 101568867
Informations de publication
Date de publication:
2020
2020
Historique:
received:
03
07
2020
accepted:
15
10
2020
entrez:
7
12
2020
pubmed:
8
12
2020
medline:
8
12
2020
Statut:
epublish
Résumé
The increased integration of molecular alterations to define tumor type or grade in central nervous system (CNS) tumor classification brings new challenges for the pathologist to make the best use of a precious limited tissue specimen for molecular studies. Within the different methods available to identify gene alterations, the droplet digital PCR (dPCR) constitutes a rapid, cost-effective, and very sensitive tool. In this study, we describe the development and validation of five multiplexed dPCR assays to detect major CNS biomarkers by using only small amounts of DNA extracted from formalin-fixed paraffin-embedded specimens. When compared to HRM-sequencing, NGS-sequencing, RNA-sequencing, or simplex digital PCR assays used as "gold standard" methods, these multiplexed dPCR assays displayed 100% specificity and sensitivity for the simultaneous detection of: 1/BRAF V600E mutation and KIAA1549:BRAF fusion; 2/FGFR1 N546K and K656E mutations and FGFR1 duplication; 3/H3F3A K27M and G34R/V mutations; 4/IDH1 R132X and IDH2 R172X mutations; and 5/TERT promoter mutations C228T and C250T. In light of the increased integration of molecular alteration, we believe that such strategies might help laboratories to optimize their screening strategies for routine diagnosis of pediatric and adult CNS tumors.
Identifiants
pubmed: 33282733
doi: 10.3389/fonc.2020.579762
pmc: PMC7689380
doi:
Types de publication
Journal Article
Langues
eng
Pagination
579762Informations de copyright
Copyright © 2020 Appay, Fina, Barets, Gallardo, Nanni-Metellus, Scavarda, Henaff, Vincent, Grewis, Pourquier, Colin and Figarella-Branger.
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