Detection of EGFR mutations in non-small cell lung cancer by droplet digital PCR.

Autor: Williamson DFK; Department of Pathology, Brigham and Women's Hospital, Boston, MA, United States of America., Marris SRN; Department of Pathology, Brigham and Women's Hospital, Boston, MA, United States of America., Rojas-Rudilla V; Department of Pathology, Brigham and Women's Hospital, Boston, MA, United States of America., Bruce JL; Department of Pathology, Brigham and Women's Hospital, Boston, MA, United States of America., Paweletz CP; Department of Medical Oncology, Dana Farber Cancer Institute, Boston, MA, United States of America., Oxnard GR; Department of Medical Oncology, Dana Farber Cancer Institute, Boston, MA, United States of America., Sholl LM; Department of Pathology, Brigham and Women's Hospital, Boston, MA, United States of America., Dong F; Department of Pathology, Brigham and Women's Hospital, Boston, MA, United States of America.
Jazyk: angličtina
Zdroj: PloS one [PLoS One] 2022 Feb 24; Vol. 17 (2), pp. e0264201. Date of Electronic Publication: 2022 Feb 24 (Print Publication: 2022).
DOI: 10.1371/journal.pone.0264201
Abstrakt: Activating mutations in EGFR predict benefit from tyrosine kinase inhibitor therapy for patients with advanced non-small cell lung cancer. Directing patients to appropriate therapy depends on accurate and timely EGFR assessment in the molecular pathology laboratory. This article describes the analytical design, performance characteristics, and clinical implementation of an assay for the rapid detection of EGFR L858R and exon 19 deletion mutations. A droplet digital polymerase chain reaction (ddPCR) assay was implemented with probe hydrolysis-dependent signal detection. A mutation-specific probe was used to detect EGFR L858R. A loss of signal design was used to detect EGFR exon 19 deletion mutations. Analytical sensitivity was dependent on DNA input and was as low as 0.01% variant allele fraction for the EGFR L858R assay and 0.1% variant allele fraction for the EGFR exon 19 deletion assay. Correlation of 20 clinical specimens tested by ddPCR and next generation sequencing showed 100% concordance. ddPCR showed 53% clinical sensitivity in the detection of EGFR mutations in plasma cell-free DNA from patients with lung cancer. The median clinical turnaround time was 5 days for ddPCR compared to 13 days for next generation sequencing. The findings show that ddPCR is an accurate and rapid method for detecting EGFR mutations in patients with non-small cell lung cancer.
Competing Interests: I have read the journal’s policy and the authors of this manuscript have the following competing interests: Geoffrey Oxnard is an employee of Foundation Medicine and holds equity in Roche. Lynette Sholl is a consultant for Genentech and Eli Lilly and receives research funding from Genentech. We can confirm that Dr. Oxnard’s and Dr. Sholl’s affiliations do not alter our adherence to PLOS ONE policies on sharing data and materials.
Databáze: MEDLINE
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