Open AccessEvaluation of two new highly multiplexed PCR assays as an alternative to next‐generation sequencing for IDH1/2 mutation detection
Author(s) -
Favre Loetitia,
Sako Nouhoum,
Tarfi Sihem,
Quang Violaine Tran,
Joy Corine,
Dupuy Aurélie,
Guillerm Erell,
Gaulard Philippe,
WagnerBallon Orianne,
Pujals Anaïs,
Sloma Ivan
Publication year - 2022
Publication title -
molecular oncology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.332
H-Index - 88
eISSN - 1878-0261
pISSN - 1574-7891
DOI - 10.1002/1878-0261.13311
Subject(s) - digital polymerase chain reaction , idh2 , idh1 , mutation , biology , cold pcr , somatic cell , dna sequencing , microbiology and biotechnology , computational biology , polymerase chain reaction , dna , point mutation , gene , genetics
IDH1 and IDH2 somatic mutations have been identified in solid tumors and blood malignancies. The development of inhibitors of mutant IDH1 and IDH2 in the past few years has prompted the development of a fast and sensitive assay to detect IDH1 R132 , IDH2 R140 and IDH2 R172 mutations to identify patients eligible for these targeted therapies. This study aimed to compare two new multiplexed PCR assays – an automated quantitative PCR (qPCR) on the PGX platform and a droplet digital PCR (ddPCR) with next‐generation sequencing (NGS) for IDH1/2 mutation detection. These assays were evaluated on 102 DNA extracted from patient peripheral blood, bone marrow and formalin‐fixed paraffin‐embedded tissue samples with mutation allelic frequency ranging from 0.6% to 45.6%. The ddPCR assay had better analytical performances than the PGX assay with 100% specificity, 100% sensitivity and a detection limit down to 0.5% on IDH1 R132 , IDH2 R140 and IDH2 R172 codons, and a high correlation with NGS results. Therefore, the new highly multiplexed ddPCR is a fast and cost‐effective assay that meets most clinical needs to identify and follow cancer patients in the era of anti‐IDH1/2‐targeted therapies.