International pilot external quality assessment scheme for analysis and reporting of circulating tumour DNA

• Published in: BMC cancer Vol. 18; no. 1; p. 804
• Main Authors: Keppens, Cleo, Dequeker, Elisabeth M C, Patton, Simon J, Normanno, Nicola, Fenizia, Francesca, Butler, Rachel, Cheetham, Melanie, Fairley, Jennifer A, Williams, Hannah, Hall, Jacqueline A, Schuuring, Ed, Deans, Zandra C
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 09-08-2018; BioMed Central; BMC
• Subjects: Biopsy; Care and treatment; cfDNA; Colorectal cancer; ctDNA; Deoxyribonucleic acid; DNA; EGFR; Gene mutation; Genetic aspects; Genetic testing; Growth factor receptors; KRAS; Laboratories; Lung cancer; Mutation; Mutation testing; Non-small cell lung carcinoma; NRAS; Pathology; Patients; Quality; Quality control; Small cell lung cancer; Tumors; United Kingdom > UK; NRAS; Lung cancer; cfDNA; Colorectal cancer; KRAS; Mutation testing; EGFR; ctDNA
• ISSN: 1471-2407; 1471-2407
• DOI: 10.1186/s12885-018-4694-x

Molecular analysis of circulating tumour DNA (ctDNA) is becoming increasingly important in clinical treatment decisions. A pilot External Quality Assessment (EQA) scheme for ctDNA analysis was organized by four European EQA providers under the umbrella organization IQN Path, in order to investigate the feasibility of delivering an EQA to assess the detection of clinically relevant variants in plasma circulating cell-free DNA (cfDNA) and to analyze reporting formats.
Thirty-two experienced laboratories received 5 samples for EGFR mutation analysis and/or 5 samples for KRAS and NRAS mutation analysis. Samples were artificially manufactured to contain 3 mL of human plasma with 20 ng/mL of fragmented ctDNA and variants at allelic frequencies of 1 and 5%.
The scheme error rate was 20.1%. Higher error rates were observed for RAS testing when compared to EGFR analysis, for allelic frequencies of 1% compared to 5%, and for cases including 2 different variants. The reports over-interpreted wild-type results and frequently failed to comment on the amount of cfDNA extracted.
The pilot scheme demonstrated the feasibility of delivering a ctDNA EQA scheme and the need for such a scheme due to high error rates in detecting low frequency clinically relevant variants. Recommendations to improve reporting of cfDNA are provided.