Evaluation of two new highly multiplexed PCR assays as an alternative to next‐generation sequencing for IDH1/2 mutation detection. Issue 22 (17th October 2022)

• Record Type: Journal Article
• Authors: Favre, Loetitia; Sako, Nouhoum; Tarfi, Sihem; Quang, Violaine Tran; Joy, Corine; Dupuy, Aurélie; Guillerm, Erell; Gaulard, Philippe; Wagner‐Ballon, Orianne; Pujals, Anaïs; Sloma, Ivan
• Is Part Of: Molecular oncology. Volume 16:Issue 22(2022)
• Page Start: 3916
• Page End: 3926
• Publication Date: 2022-10-17
• Subjects: cancer -- ddPCR -- IDH1/2 -- leukemia -- NGS -- qPCR; Cancer -- Molecular aspects -- Periodicals; 616.994005
• DOI: 10.1002/1878-0261.13311
• Languages: English
• ISSNs: 1574-7891

Abstract

Abstract : 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. Abstract : This study presents the first evaluation of two new highly multiplexed PCR assays as compared with next‐generation sequencing (NGS): an automated qPCR on the PGX platform and an allele‐specific droplet digital PCR (ddPCR) assay, both designed to detect most IDH1 and IDH2 hotspot mutations. The ddPCR assay was the best assay in terms of sensitivity, specificity, robustness, cost and timeliness, regardless of patient sample processing. It is, therefore, an appealing alternative to NGS to screen patients eligible for IDH1/2 targeted therapies.