Development and interlaboratory evaluation of a NIST Reference Material RM 8366 for EGFR and MET gene copy number measurements.

Autor: He HJ; Biosystems and Biomaterials Division, National Institute of Standards and Technology, 100 Bureau Drive, MS 8312, Gaithersburg, MD 20899, USA, Phone: +301-975-2169, Fax: +301-330-3447., Das B; Molecular Characterization and Clinical Assay Development Laboratory, Frederick National Laboratory for Cancer Research, Frederick, MD, USA., Cleveland MH; Biomolecular Measurement Division, National Institute of Standards and Technology, Gaithersburg, MD, USA., Chen L; Molecular Characterization and Clinical Assay Development Laboratory, Frederick National Laboratory for Cancer Research, Frederick, MD, USA., Camalier CE; Molecular Characterization and Clinical Assay Development Laboratory, Frederick National Laboratory for Cancer Research, Frederick, MD, USA., Liu LC; Thermo Fisher Scientific, Fremont, CA, USA., Norman KL; Thermo Fisher Scientific, Fremont, CA, USA., Fellowes AP; Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia., McEvoy CR; Peter MacCallum Cancer Centre, Melbourne, Victoria, Australia., Lund SP; Statistical Engineering Division, National Institute of Standards and Technology, Gaithersburg, MD, USA., Almeida J; Biosystems and Biomaterials Division, National Institute of Standards and Technology, Gaithersburg, MD, USA., Steffen CR; Biomolecular Measurement Division, National Institute of Standards and Technology, Gaithersburg, MD, USA., Karlovich C; Molecular Characterization and Clinical Assay Development Laboratory, Frederick National Laboratory for Cancer Research, Frederick, MD, USA., Williams PM; Molecular Characterization and Clinical Assay Development Laboratory, Frederick National Laboratory for Cancer Research, Frederick, MD, USA., Cole KD; Biosystems and Biomaterials Division, National Institute of Standards and Technology, 100 Bureau Drive, MS 8312, Gaithersburg, MD 20899, USA, Phone: +301-975-2169, Fax: +301-330-3447.
Jazyk: angličtina
Zdroj: Clinical chemistry and laboratory medicine [Clin Chem Lab Med] 2019 Jul 26; Vol. 57 (8), pp. 1142-1152.
DOI: 10.1515/cclm-2018-1306
Abstrakt: Background The National Institute of Standards and Technology (NIST) Reference Material RM 8366 was developed to improve the quality of gene copy measurements of EGFR (epidermal growth factor receptor) and MET (proto-oncogene, receptor tyrosine kinase), important targets for cancer diagnostics and treatment. The reference material is composed of genomic DNA prepared from six human cancer cell lines with different levels of amplification of the target genes. Methods The reference values for the ratios of the EGFR and MET gene copy numbers to the copy numbers of reference genes were measured using digital PCR. The digital PCR measurements were confirmed by two additional laboratories. The samples were also characterized using Next Generation Sequencing (NGS) methods including whole genome sequencing (WGS) at three levels of coverage (approximately 1 ×, 5 ×  and greater than 30 ×), whole exome sequencing (WES), and two different pan-cancer gene panels. The WES data were analyzed using three different bioinformatic algorithms. Results The certified values (digital PCR) for EGFR and MET were in good agreement (within 20%) with the values obtained from the different NGS methods and algorithms for five of the six components; one component had lower NGS values. Conclusions This study shows that NIST RM 8366 is a valuable reference material to evaluate the performance of assays that assess EGFR and MET gene copy number measurements.
Databáze: MEDLINE
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