title: Initial Diagnosis of ALK-Positive Non-Small-Cell Lung Cancer Based on Analysis of ALK Status Utilizing Droplet Digital PCR.
abstract: We describe a novel droplet digital PCR (ddPCR) assay capable of detecting genomic alterations associated with inversion translocations. It is applied here to detection of rearrangements in the anaplastic lymphoma kinase (ALK) gene associated with ALK-positive non-small-cell lung cancer (NSCLC). NSCLC patients may carry a nonreciprocal translocation on human chromosome 2, in which synchronized double stranded breaks (DSB) within the echinoderm microtubule-associated protein-like 4 (EML4) gene and ALK lead to an inversion of genetic material that forms the non-natural gene fusion EML4-ALK encoding a constitutively active tyrosine kinase that is associated with 3 to 7% of all NSCLCs. Detection of ALK rearrangements is currently achieved in clinics through direct visualization via a fluorescent in situ hybridization (FISH) assay, which can detect those rearrangements to a limit of detection (LOD) of ca. 15%. We show that the ddPCR assay presented here provides a LOD of 0.25% at lower cost and with faster turnaround times.
authors.affiliation: Michael Smith Laboratories, University of British Columbia , Vancouver, British Columbia V6T 1Z4, Canada.|Genomic Science and Technology, University of British Columbia , Vancouver, British Columbia V5Z 4S6, Canada.
authors.forename: H Louise
authors.initials: HL
authors.lastname: Lund
authors.affiliation: Michael Smith Laboratories, University of British Columbia , Vancouver, British Columbia V6T 1Z4, Canada.|Department of Chemical and Biological Engineering, University of British Columbia , Vancouver, British Columbia V6T 1Z3, Canada.
authors.forename: Curtis B
authors.initials: CB
authors.lastname: Hughesman
authors.affiliation: Michael Smith Laboratories, University of British Columbia , Vancouver, British Columbia V6T 1Z4, Canada.|Department of Chemical and Biological Engineering, University of British Columbia , Vancouver, British Columbia V6T 1Z3, Canada.
authors.forename: Kareem
authors.initials: K
authors.lastname: Fakhfakh
authors.affiliation: Cancer Genetics Testing Laboratory, Department of Pathology and Laboratory Medicine, British Columbia Cancer Agency, Vancouver, British Columbia V5Z 4E6, Canada.
authors.forename: Kelly
authors.initials: K
authors.lastname: McNeil
authors.affiliation: Cancer Genetics Testing Laboratory, Department of Pathology and Laboratory Medicine, British Columbia Cancer Agency, Vancouver, British Columbia V5Z 4E6, Canada.
authors.forename: Shahira
authors.initials: S
authors.lastname: Clemens
authors.affiliation: Cancer Genetics Testing Laboratory, Department of Pathology and Laboratory Medicine, British Columbia Cancer Agency, Vancouver, British Columbia V5Z 4E6, Canada.
authors.forename: Kimberly
authors.initials: K
authors.lastname: Hocken
authors.affiliation: Cancer Genetics Testing Laboratory, Department of Pathology and Laboratory Medicine, British Columbia Cancer Agency, Vancouver, British Columbia V5Z 4E6, Canada.
authors.forename: Ryan
authors.initials: R
authors.lastname: Pettersson
authors.affiliation: Cancer Genetics Testing Laboratory, Department of Pathology and Laboratory Medicine, British Columbia Cancer Agency, Vancouver, British Columbia V5Z 4E6, Canada.
authors.forename: Aly
authors.initials: A
authors.lastname: Karsan
authors.affiliation: Michael Smith Laboratories, University of British Columbia , Vancouver, British Columbia V6T 1Z4, Canada.|Department of Biochemistry and Molecular Biology, University of British Columbia , Vancouver, British Columbia V6T 1Z3, Canada.
authors.forename: Leonard J
authors.initials: LJ
authors.lastname: Foster
authors.affiliation: Michael Smith Laboratories, University of British Columbia , Vancouver, British Columbia V6T 1Z4, Canada.|Genomic Science and Technology, University of British Columbia , Vancouver, British Columbia V5Z 4S6, Canada.|Department of Chemical and Biological Engineering, University of British Columbia , Vancouver, British Columbia V6T 1Z3, Canada.
authors.forename: Charles
authors.initials: C
authors.lastname: Haynes
mesh_terms: D000077548:Anaplastic Lymphoma Kinase; D002289:Carcinoma, Non-Small-Cell Lung / Q000175:diagnosis* / Q000378:metabolism; D006801:Humans; D008175:Lung Neoplasms / Q000175:diagnosis* / Q000378:metabolism; D010316:Particle Size; D016133:Polymerase Chain Reaction / Q000379:methods*; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics* / Q000378:metabolism
chemical_list: C000626173:ALK protein, human; D000077548:Anaplastic Lymphoma Kinase; D020794:Receptor Protein-Tyrosine Kinases