Establishment of primary reference measurement procedures and reference materials for EGFR variant detection in non-small cell lung cancer
| Autor: | Lianhua Dong, Lan Qingkuo, Yongzhuo Zhang, Pengyu Zhu, Chunyan Niu, Lingxiang Zhu, Shangjun Wang, Wentao Xu, Gao Huafang, Guo Yong, Yunhua Gao, Xiaoyan Cheng, Meihong Du, Xiaohua Jin, Li Liang, Xia Wang |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
Oncology
0303 health sciences medicine.medical_specialty Reproducibility business.industry General Chemical Engineering Coefficient of variation General Engineering Cancer Repeatability medicine.disease Analytical Chemistry 03 medical and health sciences T790M 0302 clinical medicine 030220 oncology & carcinogenesis Internal medicine Cancer screening medicine Digital polymerase chain reaction business Genotyping 030304 developmental biology |
| Zdroj: | Analytical Methods. 13:2114-2123 |
| ISSN: | 1759-9679 1759-9660 |
| DOI: | 10.1039/d1ay00328c |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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