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dc.contributor.authorPender, Aen_US
dc.contributor.authorGarcia-Murillas, Ien_US
dc.contributor.authorRana, Sen_US
dc.contributor.authorCutts, RJen_US
dc.contributor.authorKelly, Gen_US
dc.contributor.authorFenwick, Ken_US
dc.contributor.authorKozarewa, Ien_US
dc.contributor.authorGonzalez de Castro, Den_US
dc.contributor.authorBhosle, Jen_US
dc.contributor.authorO'Brien, Men_US
dc.contributor.authorTurner, NCen_US
dc.contributor.authorPopat, Sen_US
dc.contributor.authorDownward, Jen_US
dc.date.accessioned2018-11-29T09:20:04Z
dc.date.issued2015-01
dc.identifier.citationPloS one, 2015, 10 (9), pp. e0139074 - ?
dc.identifier.issn1932-6203
dc.identifier.urihttps://repository.icr.ac.uk/handle/internal/2962
dc.identifier.eissn1932-6203
dc.identifier.doi10.1371/journal.pone.0139074
dc.description.abstractDroplet digital PCR (ddPCR) can be used to detect low frequency mutations in oncogene-driven lung cancer. The range of KRAS point mutations observed in NSCLC necessitates a multiplex approach to efficient mutation detection in circulating DNA. Here we report the design and optimisation of three discriminatory ddPCR multiplex assays investigating nine different KRAS mutations using PrimePCR™ ddPCR™ Mutation Assays and the Bio-Rad QX100 system. Together these mutations account for 95% of the nucleotide changes found in KRAS in human cancer. Multiplex reactions were optimised on genomic DNA extracted from KRAS mutant cell lines and tested on DNA extracted from fixed tumour tissue from a cohort of lung cancer patients without prior knowledge of the specific KRAS genotype. The multiplex ddPCR assays had a limit of detection of better than 1 mutant KRAS molecule in 2,000 wild-type KRAS molecules, which compared favourably with a limit of detection of 1 in 50 for next generation sequencing and 1 in 10 for Sanger sequencing. Multiplex ddPCR assays thus provide a highly efficient methodology to identify KRAS mutations in lung adenocarcinoma.
dc.formatElectronic-eCollection
dc.format.extente0139074 - ?
dc.languageeng
dc.language.isoeng
dc.rights.urihttps://creativecommons.org/licenses/by/4.0
dc.subjectCell Line, Tumor
dc.subjectClone Cells
dc.subjectHumans
dc.subjectCarcinoma, Non-Small-Cell Lung
dc.subjectLung Neoplasms
dc.subjectFormaldehyde
dc.subjectDNA, Neoplasm
dc.subjectParaffin Embedding
dc.subjectTissue Fixation
dc.subjectTemperature
dc.subjectGene Frequency
dc.subjectMutation
dc.subjectProto-Oncogene Proteins p21(ras)
dc.subjectHigh-Throughput Nucleotide Sequencing
dc.subjectGenotyping Techniques
dc.subjectMultiplex Polymerase Chain Reaction
dc.titleEfficient Genotyping of KRAS Mutant Non-Small Cell Lung Cancer Using a Multiplexed Droplet Digital PCR Approach.
dc.typeJournal Article
dcterms.dateAccepted2015-09-09
rioxxterms.versionofrecord10.1371/journal.pone.0139074
rioxxterms.licenseref.urihttps://creativecommons.org/licenses/by/4.0
rioxxterms.licenseref.startdate2015-01
rioxxterms.typeJournal Article/Review
dc.relation.isPartOfPloS one
pubs.issue9
pubs.notesNot known
pubs.organisational-group/ICR
pubs.organisational-group/ICR/Primary Group
pubs.organisational-group/ICR/Primary Group/ICR Divisions
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Breast Cancer Research
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Breast Cancer Research/Molecular Oncology
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Cancer Biology
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Cancer Biology/Lung Cancer Group
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Clinical Studies
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Clinical Studies/Thoracic Oncology
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Clinical Studies/Thoracic Oncology/Thoracic Oncology (hon.)
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Clinical Studies/Treatment of thoracic tumours
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Clinical Studies/Treatment of thoracic tumours/Treatment of thoracic tumours (hon.)
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Molecular Pathology
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Molecular Pathology/Lung Cancer Group
pubs.organisational-group/ICR/Primary Group/Royal Marsden Clinical Units
pubs.publication-statusPublished
pubs.volume10
pubs.embargo.termsNot known
icr.researchteamMolecular Oncologyen_US
icr.researchteamThoracic Oncologyen_US
icr.researchteamTreatment of thoracic tumoursen_US
icr.researchteamLung Cancer Groupen_US
dc.contributor.icrauthorDownward, Julian David Harryen
dc.contributor.icrauthorGonzalez de Castro, Daviden
dc.contributor.icrauthorO'Brien, Maryen
dc.contributor.icrauthorTurner, Nicholasen
dc.contributor.icrauthorPopat, Sanjayen
dc.contributor.icrauthorGarcia-Murillas, Isaacen
dc.contributor.icrauthorPender, Alexandraen


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