A Pan-Canadian Validation Study for the Detection of EGFR T790M Mutation Using Circulating Tumor DNA From Peripheral Blood
Introduction: Genotyping circulating tumor DNA (ctDNA) is a promising noninvasive clinical tool to identify the EGFR T790M resistance mutation in patients with advanced NSCLC with resistance to EGFR inhibitors. To facilitate standardization and clinical adoption of ctDNA testing across Canada, we de...
| Main Authors: | , , , , , , , , , , , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2021-08-01
|
| Series: | JTO Clinical and Research Reports |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666364321000710 |
| _version_ | 1818366735353380864 |
|---|---|
| author | Shamini Selvarajah, PhD Sophie Plante, MSc Marsha Speevak, PhD Andrea Vaags, PhD Darren Hamelinck, MSc Martin Butcher, MSc Elizabeth McCready, PhD Daria Grafodatskaya, PhD Normand Blais, MD Danh Tran-Thanh, MD Xiaoduan Weng, MD Rami Nassabein, MD Wenda Greer, PhD Ryan N. Walton, MPH Bryan Lo, MD Doug Demetrick, MD Stephanie Santos, BSc Bekim Sadikovic, PhD Xiao Zhang, BSc, MSc Tong Zhang, MD Tara Spence, PhD Tracy Stockley, PhD Harriet Feilotter, PhD Philippe Joubert, MD, PhD |
| author_facet | Shamini Selvarajah, PhD Sophie Plante, MSc Marsha Speevak, PhD Andrea Vaags, PhD Darren Hamelinck, MSc Martin Butcher, MSc Elizabeth McCready, PhD Daria Grafodatskaya, PhD Normand Blais, MD Danh Tran-Thanh, MD Xiaoduan Weng, MD Rami Nassabein, MD Wenda Greer, PhD Ryan N. Walton, MPH Bryan Lo, MD Doug Demetrick, MD Stephanie Santos, BSc Bekim Sadikovic, PhD Xiao Zhang, BSc, MSc Tong Zhang, MD Tara Spence, PhD Tracy Stockley, PhD Harriet Feilotter, PhD Philippe Joubert, MD, PhD |
| author_sort | Shamini Selvarajah, PhD |
| collection | DOAJ |
| description | Introduction: Genotyping circulating tumor DNA (ctDNA) is a promising noninvasive clinical tool to identify the EGFR T790M resistance mutation in patients with advanced NSCLC with resistance to EGFR inhibitors. To facilitate standardization and clinical adoption of ctDNA testing across Canada, we developed a 2-phase multicenter study to standardize T790M mutation detection using plasma ctDNA testing. Methods: In phase 1, commercial reference standards were distributed to participating clinical laboratories, to use their existing platforms for mutation detection. Baseline performance characteristics were established using known and blinded engineered plasma samples spiked with predetermined concentrations of T790M, L858R, and exon 19 deletion variants. In phase II, peripheral blood collected from local patients with known EGFR activating mutations and progressing on treatment were assayed for the presence of EGFR variants and concordance with a clinically validated test at the reference laboratory. Results: All laboratories in phase 1 detected the variants at 0.5 % and 5.0 % allele frequencies, with no false positives. In phase 2, the concordance with the reference laboratory for detection of both the primary and resistance mutation was high, with next-generation sequencing and droplet digital polymerase chain reaction exhibiting the best overall concordance. Data also suggested that the ability to detect mutations at clinically relevant limits of detection is generally not platform-specific, but rather impacted by laboratory-specific practices. Conclusions: Discrepancies among sending laboratories using the same assay suggest that laboratory-specific practices may impact performance. In addition, a negative or inconclusive ctDNA test should be followed by tumor testing when possible. |
| first_indexed | 2024-12-13T22:40:53Z |
| format | Article |
| id | doaj.art-4343799e3fbf40999e9a88df77f2790e |
| institution | Directory Open Access Journal |
| issn | 2666-3643 |
| language | English |
| last_indexed | 2024-12-13T22:40:53Z |
| publishDate | 2021-08-01 |
| publisher | Elsevier |
| record_format | Article |
| series | JTO Clinical and Research Reports |
| spelling | doaj.art-4343799e3fbf40999e9a88df77f2790e2022-12-21T23:28:52ZengElsevierJTO Clinical and Research Reports2666-36432021-08-0128100212A Pan-Canadian Validation Study for the Detection of EGFR T790M Mutation Using Circulating Tumor DNA From Peripheral BloodShamini Selvarajah, PhD0Sophie Plante, MSc1Marsha Speevak, PhD2Andrea Vaags, PhD3Darren Hamelinck, MSc4Martin Butcher, MSc5Elizabeth McCready, PhD6Daria Grafodatskaya, PhD7Normand Blais, MD8Danh Tran-Thanh, MD9Xiaoduan Weng, MD10Rami Nassabein, MD11Wenda Greer, PhD12Ryan N. Walton, MPH13Bryan Lo, MD14Doug Demetrick, MD15Stephanie Santos, BSc16Bekim Sadikovic, PhD17Xiao Zhang, BSc, MSc18Tong Zhang, MD19Tara Spence, PhD20Tracy Stockley, PhD21Harriet Feilotter, PhD22Philippe Joubert, MD, PhD23Department of Laboratory Medicine and Genetics, Trillium Health Partners, Mississauga, Ontario, Canada; Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Ontario, CanadaInstitut Universitaire de Cardiologie et de Pneumologie de Québec-Université Laval, Québec, Quebec, CanadaDepartment of Laboratory Medicine and Genetics, Trillium Health Partners, Mississauga, Ontario, Canada; Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Ontario, CanadaDepartment of Laboratory Medicine and Genetics, Trillium Health Partners, Mississauga, Ontario, Canada; Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Ontario, CanadaDepartment of Laboratory Medicine and Genetics, Trillium Health Partners, Mississauga, Ontario, CanadaDepartment of Oncology, McMaster University, Hamilton, Ontario, CanadaDepartment of Pathology and Molecular Medicine, McMaster University, Hamilton, Ontario, Canada; Hamilton Regional Laboratory Medicine Program, Hamilton Health Sciences, Hamilton, Ontario, CanadaDepartment of Pathology and Molecular Medicine, McMaster University, Hamilton, Ontario, Canada; Hamilton Regional Laboratory Medicine Program, Hamilton Health Sciences, Hamilton, Ontario, CanadaCentre Hospitalier de l’Université de Montréal, Montréal, Quebec, CanadaCentre Hospitalier de l’Université de Montréal, Montréal, Quebec, CanadaCentre Hospitalier de l’Université de Montréal, Montréal, Quebec, CanadaCentre Hospitalier de l’Université de Montréal, Montréal, Quebec, CanadaQueen Elizabeth II Health Sciences Center, Department of Pathology, Faculty of Medicine, Dalhousie University, Halifax, Nova Scotia, CanadaAstraZeneca Canada, Mississauga, Ontario, CanadaDepartment of Pathology and Laboratory Medicine, The Ottawa Hospital, Ottawa, Ontario, CanadaDepartment of Pathology and Laboratory Medicine, University of Calgary, Calgary, Alberta, Canada; Department of Oncology, University of Calgary, Calgary, Alberta, Canada; Department of Biochemistry and Molecular Biology, University of Calgary, Calgary, Alberta, Canada; Department of Medical Genetics, University of Calgary, Calgary, Alberta, CanadaMolecular Genetics Laboratory, Molecular Diagnostics Division, London Health Sciences Centre, London, Ontario, CanadaMolecular Genetics Laboratory, Molecular Diagnostics Division, London Health Sciences Centre, London, Ontario, Canada; Department of Pathology and Laboratory Medicine, Western University, London, Ontario, Canada; Verspeeten Clinical Genome Centre, London Health Sciences Centre, London, Ontario, CanadaLaboratory Genetics, Kingston Health Sciences Center, Kingston, Ontario, CanadaDepartment of Clinical Laboratory Genetics, University Health Network, Toronto, Ontario, CanadaDepartment of Pathology and Molecular Medicine, Queen’s University, Kingston, Ontario, CanadaDepartment of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Ontario, Canada; Department of Clinical Laboratory Genetics, University Health Network, Toronto, Ontario, CanadaLaboratory Genetics, Kingston Health Sciences Center, Kingston, Ontario, Canada; Department of Pathology and Molecular Medicine, Queen’s University, Kingston, Ontario, CanadaInstitut Universitaire de Cardiologie et de Pneumologie de Québec-Université Laval, Québec, Quebec, Canada; Corresponding author. Address for correspondence: Philippe Joubert, MD, PhD, Institut Universitaire de Cardiologie et de Pneumologie de Québec-Université Laval, 2725 chemin Sainte-Foy, C2259, Québec, QC, Canada, G1V4G5.Introduction: Genotyping circulating tumor DNA (ctDNA) is a promising noninvasive clinical tool to identify the EGFR T790M resistance mutation in patients with advanced NSCLC with resistance to EGFR inhibitors. To facilitate standardization and clinical adoption of ctDNA testing across Canada, we developed a 2-phase multicenter study to standardize T790M mutation detection using plasma ctDNA testing. Methods: In phase 1, commercial reference standards were distributed to participating clinical laboratories, to use their existing platforms for mutation detection. Baseline performance characteristics were established using known and blinded engineered plasma samples spiked with predetermined concentrations of T790M, L858R, and exon 19 deletion variants. In phase II, peripheral blood collected from local patients with known EGFR activating mutations and progressing on treatment were assayed for the presence of EGFR variants and concordance with a clinically validated test at the reference laboratory. Results: All laboratories in phase 1 detected the variants at 0.5 % and 5.0 % allele frequencies, with no false positives. In phase 2, the concordance with the reference laboratory for detection of both the primary and resistance mutation was high, with next-generation sequencing and droplet digital polymerase chain reaction exhibiting the best overall concordance. Data also suggested that the ability to detect mutations at clinically relevant limits of detection is generally not platform-specific, but rather impacted by laboratory-specific practices. Conclusions: Discrepancies among sending laboratories using the same assay suggest that laboratory-specific practices may impact performance. In addition, a negative or inconclusive ctDNA test should be followed by tumor testing when possible.http://www.sciencedirect.com/science/article/pii/S2666364321000710Non–small cell lung cancerPlasma ctDNA testingLiquid biopsyEGFR T790M variant |
| spellingShingle | Shamini Selvarajah, PhD Sophie Plante, MSc Marsha Speevak, PhD Andrea Vaags, PhD Darren Hamelinck, MSc Martin Butcher, MSc Elizabeth McCready, PhD Daria Grafodatskaya, PhD Normand Blais, MD Danh Tran-Thanh, MD Xiaoduan Weng, MD Rami Nassabein, MD Wenda Greer, PhD Ryan N. Walton, MPH Bryan Lo, MD Doug Demetrick, MD Stephanie Santos, BSc Bekim Sadikovic, PhD Xiao Zhang, BSc, MSc Tong Zhang, MD Tara Spence, PhD Tracy Stockley, PhD Harriet Feilotter, PhD Philippe Joubert, MD, PhD A Pan-Canadian Validation Study for the Detection of EGFR T790M Mutation Using Circulating Tumor DNA From Peripheral Blood JTO Clinical and Research Reports Non–small cell lung cancer Plasma ctDNA testing Liquid biopsy EGFR T790M variant |
| title | A Pan-Canadian Validation Study for the Detection of EGFR T790M Mutation Using Circulating Tumor DNA From Peripheral Blood |
| title_full | A Pan-Canadian Validation Study for the Detection of EGFR T790M Mutation Using Circulating Tumor DNA From Peripheral Blood |
| title_fullStr | A Pan-Canadian Validation Study for the Detection of EGFR T790M Mutation Using Circulating Tumor DNA From Peripheral Blood |
| title_full_unstemmed | A Pan-Canadian Validation Study for the Detection of EGFR T790M Mutation Using Circulating Tumor DNA From Peripheral Blood |
| title_short | A Pan-Canadian Validation Study for the Detection of EGFR T790M Mutation Using Circulating Tumor DNA From Peripheral Blood |
| title_sort | pan canadian validation study for the detection of egfr t790m mutation using circulating tumor dna from peripheral blood |
| topic | Non–small cell lung cancer Plasma ctDNA testing Liquid biopsy EGFR T790M variant |
| url | http://www.sciencedirect.com/science/article/pii/S2666364321000710 |
| work_keys_str_mv | AT shaminiselvarajahphd apancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT sophieplantemsc apancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT marshaspeevakphd apancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT andreavaagsphd apancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT darrenhamelinckmsc apancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT martinbutchermsc apancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT elizabethmccreadyphd apancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT dariagrafodatskayaphd apancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT normandblaismd apancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT danhtranthanhmd apancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT xiaoduanwengmd apancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT raminassabeinmd apancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT wendagreerphd apancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT ryannwaltonmph apancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT bryanlomd apancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT dougdemetrickmd apancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT stephaniesantosbsc apancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT bekimsadikovicphd apancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT xiaozhangbscmsc apancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT tongzhangmd apancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT taraspencephd apancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT tracystockleyphd apancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT harrietfeilotterphd apancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT philippejoubertmdphd apancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT shaminiselvarajahphd pancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT sophieplantemsc pancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT marshaspeevakphd pancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT andreavaagsphd pancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT darrenhamelinckmsc pancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT martinbutchermsc pancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT elizabethmccreadyphd pancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT dariagrafodatskayaphd pancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT normandblaismd pancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT danhtranthanhmd pancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT xiaoduanwengmd pancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT raminassabeinmd pancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT wendagreerphd pancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT ryannwaltonmph pancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT bryanlomd pancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT dougdemetrickmd pancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT stephaniesantosbsc pancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT bekimsadikovicphd pancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT xiaozhangbscmsc pancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT tongzhangmd pancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT taraspencephd pancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT tracystockleyphd pancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT harrietfeilotterphd pancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood AT philippejoubertmdphd pancanadianvalidationstudyforthedetectionofegfrt790mmutationusingcirculatingtumordnafromperipheralblood |