Jump to main content
Jump to site search

Issue 18, 2021
Previous Article Next Article

Establishment of primary reference measurement procedures and reference materials for EGFR variant detection in non-small cell lung cancer

Author affiliations

Abstract

Circulating tumor DNA (ctDNA)-based mutation detection is promising to change the clinical practice of genotype-directed therapy for cancer. A growing number of non-invasive tests for cancer screening and monitoring that involve the detection of ctDNA have been commercialized. Primary reference measurement procedures (PRMPs) and reference materials (RMs) are urgently needed to assess the non-invasive tests. In this study, a PRMP based on digital PCR (dPCR) and ctDNA RMs for quantification of the frequently occurring variant in epidermal growth factor receptor (EGFR L858R, T790M, and 19Del) in non-small cell lung cancer (NSCLC) were established. The candidate dPCR PRMP showed high specificity (false positive rate 0–0.003%), good repeatability (coefficient of variance (CV), 2–3% for 104 copies/reaction), and high interlaboratory reproducibility (3–10%). A good linearity (0.97 < slope < 1.03, R2 ≥ 0.9999) between the measured mutant (MU) value and prepared value was observed for all assays over the fractional abundance (FA) range, between 25% and 0.05%. The limit of quantification (LoQ) was determined to be 34 L858R, 23 T790M, and 34 19Del copies/reaction, corresponding to a FA of 0.2%. An inter-laboratory study of using the EGFR ctDNA RMs and dPCR assays demonstrated that the participating laboratories produced consistent concentrations of MU and wild-type (WT), as well as FA. This study demonstrates that dPCR can act as a potential PRMP for EGFR mutation for validation of NSCLC genotyping tests and ctDNA quantitative tests. The PRMP and RMs established here could improve interlaboratory repeatability and reproducibility, which supports rapid translation and application of non-invasive tests into clinical practice.

Graphical abstract: Establishment of primary reference measurement procedures and reference materials for EGFR variant detection in non-small cell lung cancer

Back to tab navigation

Supplementary files

Article information


Submitted
25 Feb 2021
Accepted
25 Mar 2021
First published
19 Apr 2021

Anal. Methods, 2021,13, 2114-2123
Article type
Paper

Establishment of primary reference measurement procedures and reference materials for EGFR variant detection in non-small cell lung cancer

X. Wang, Y. Zhang, C. Niu, S. Wang, L. Li, Y. Guo, L. Zhu, X. Jin, H. Gao, W. Xu, P. Zhu, Q. Lan, M. Du, X. Cheng, Y. Gao and L. Dong, Anal. Methods, 2021, 13, 2114
DOI: 10.1039/D1AY00328C

Social activity

Search articles by author

Spotlight

Advertisements