International Interlaboratory Digital PCR Study Demonstrating High Reproducibility for the Measurement of a Rare Sequence Variant.

Autor: Whale AS; Molecular and Cell Biology Team, LGC , Queens Road, Teddington, Middlesex TW11 0LY, United Kingdom., Devonshire AS; Molecular and Cell Biology Team, LGC , Queens Road, Teddington, Middlesex TW11 0LY, United Kingdom., Karlin-Neumann G; Digital Biology Center, Bio-Rad Laboratories , 5731 West Las Positas, Pleasanton, California 94588, United States., Regan J; Digital Biology Center, Bio-Rad Laboratories , 5731 West Las Positas, Pleasanton, California 94588, United States., Javier L; Digital Biology Center, Bio-Rad Laboratories , 5731 West Las Positas, Pleasanton, California 94588, United States., Cowen S; Statistics Team, LGC , Queens Road, Teddington, Middlesex TW11 0LY, United Kingdom., Fernandez-Gonzalez A; Molecular and Cell Biology Team, LGC , Queens Road, Teddington, Middlesex TW11 0LY, United Kingdom., Jones GM; Molecular and Cell Biology Team, LGC , Queens Road, Teddington, Middlesex TW11 0LY, United Kingdom., Redshaw N; Molecular and Cell Biology Team, LGC , Queens Road, Teddington, Middlesex TW11 0LY, United Kingdom., Beck J; Chronix Biomedical , Goetheallee 8, 37073 Goettingen, Germany., Berger AW; Department of Internal Medicine I, Ulm University , Albert-Einstein-Allee 23, 89081, Ulm, Germany., Combaret V; Laboratoire de Recherche Translationnelle, Centre Léon-Bérard , Lyon, F-69008, France., Dahl Kjersgaard N; Sjællands Universitetshospital , Klinisk Biokemisk Afdeling, Molekylærenhed, Sygehusvej 10, 4000 Roskilde, Denmark., Davis L; Genoptix Inc. , 1811 Aston Avenue, Carlsbad, California 92008, United States., Fina F; Service de Transfert d'Oncologie Biologique, Laboratoire de Biologie Médicale, Faculte de médecine Nord , Boulevard Pierre Dramard, Marseille 13916 cedex 20, France., Forshew T; UCL Cancer Institute , Paul O'Gorman Building, 72 Huntley Street, London WC1E 6DD, United Kingdom., Fredslund Andersen R; Department of Clinical Immunology and Biochemistry, Vejle Hospital , Kabbeltoft 25, 7100 Vejle, Denmark., Galbiati S; Division of Genetics and Cell Biology, IRCCS San Raffaele Scientific Institute , via Olgettina 60, 20132 Milano, Italy., González Hernández Á; Department of Pediatrics and CIMA LAB Diagnostics, Clínica Universidad de Navarra , Avenida Pío XII 36, 31008 Pamplona, Spain., Haynes CA; Michael Smith Laboratories, University of British Columbia , 301 Michael Smith Building, 2185 East Mall, Vancouver, BC V6T 1Z4, Canada., Janku F; The University of Texas MD Anderson Cancer Center , Department of Investigational Cancer Therapeutics (Phase I Clinical Trials Program), 1515 Holcombe Boulevard 0455, Houston, Texas 77030, United States., Lacave R; Solid Tumours Genomics Unit, Tenon Hospital, APHP and Université Pierre et Marie Curie , 4 rue de la Chine, 75970 Paris, France., Lee J; Johns Hopkins , 1650 Orleans St., Baltimore, Maryland 21287, United States., Mistry V; Department of Cancer Studies, University of Leicester , Robert Kilpatrick Clinical Sciences Building, Leicester Royal Infirmary, Leicester LE2 7LX, United Kingdom., Pender A; Lung Cancer Group, Division of Molecular Pathology, The Institute of Cancer Research , 123 Old Brompton Road, London SW7 3RP, United Kingdom., Pradines A; Institut Claudius Regaud - IUCTO , Laboratoire de Biologie Medicale Oncologique, 1 avenue Irène Joliot-Curie, Toulouse 31059 cedex 9, France., Proudhon C; Institut Curie, PSL Research University, SiRIC , Laboratory of Circulating Tumor Biomarkers, 26 rue d'Ulm, 75005 Paris, France., Saal LH; Department of Oncology and Pathology, Lund University , Scheelevägen 2, MV 404-B2, SE-22381, Lund, Sweden., Stieglitz E; Department of Pediatrics, Benioff Children's Hospital, UCSF Helen Diller Family Comprehensive Cancer Center , 1450 Third Street, San Francisco, California 94158, United States., Ulrich B; Dana Farber Cancer Institute , Belfer Center for Applied Cancer Science and Department of Medical Oncology, Boston, Massachusetts 02115, United States., Foy CA; Molecular and Cell Biology Team, LGC , Queens Road, Teddington, Middlesex TW11 0LY, United Kingdom., Parkes H; Molecular and Cell Biology Team, LGC , Queens Road, Teddington, Middlesex TW11 0LY, United Kingdom., Tzonev S; Digital Biology Center, Bio-Rad Laboratories , 5731 West Las Positas, Pleasanton, California 94588, United States., Huggett JF; Molecular and Cell Biology Team, LGC , Queens Road, Teddington, Middlesex TW11 0LY, United Kingdom.; School of Biosciences & Medicine, Faculty of Health & Medical Science, University of Surrey , Guildford, Surrey GU2 7XH, United Kingdom.
Jazyk: angličtina
Zdroj: Analytical chemistry [Anal Chem] 2017 Feb 07; Vol. 89 (3), pp. 1724-1733. Date of Electronic Publication: 2017 Jan 18.
DOI: 10.1021/acs.analchem.6b03980
Abstrakt: This study tested the claim that digital PCR (dPCR) can offer highly reproducible quantitative measurements in disparate laboratories. Twenty-one laboratories measured four blinded samples containing different quantities of a KRAS fragment encoding G12D, an important genetic marker for guiding therapy of certain cancers. This marker is challenging to quantify reproducibly using quantitative PCR (qPCR) or next generation sequencing (NGS) due to the presence of competing wild type sequences and the need for calibration. Using dPCR, 18 laboratories were able to quantify the G12D marker within 12% of each other in all samples. Three laboratories appeared to measure consistently outlying results; however, proper application of a follow-up analysis recommendation rectified their data. Our findings show that dPCR has demonstrable reproducibility across a large number of laboratories without calibration. This could enable the reproducible application of molecular stratification to guide therapy and, potentially, for molecular diagnostics.
Databáze: MEDLINE