dc.contributor.author | Pender, A | |
dc.contributor.author | Garcia-Murillas, I | |
dc.contributor.author | Rana, S | |
dc.contributor.author | Cutts, RJ | |
dc.contributor.author | Kelly, G | |
dc.contributor.author | Fenwick, K | |
dc.contributor.author | Kozarewa, I | |
dc.contributor.author | Gonzalez de Castro, D | |
dc.contributor.author | Bhosle, J | |
dc.contributor.author | O'Brien, M | |
dc.contributor.author | Turner, NC | |
dc.contributor.author | Popat, S | |
dc.contributor.author | Downward, J | |
dc.date.accessioned | 2018-11-29T09:20:04Z | |
dc.date.issued | 2015-09-28 | |
dc.identifier.citation | PloS one, 2015, 10 (9), pp. e0139074 - ? | |
dc.identifier.issn | 1932-6203 | |
dc.identifier.uri | https://repository.icr.ac.uk/handle/internal/2962 | |
dc.identifier.eissn | 1932-6203 | |
dc.identifier.doi | 10.1371/journal.pone.0139074 | |
dc.description.abstract | Droplet 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.format | Electronic-eCollection | |
dc.format.extent | e0139074 - ? | |
dc.language | eng | |
dc.language.iso | eng | |
dc.publisher | PUBLIC LIBRARY SCIENCE | |
dc.rights.uri | https://creativecommons.org/licenses/by/4.0 | |
dc.subject | Cell Line, Tumor | |
dc.subject | Clone Cells | |
dc.subject | Humans | |
dc.subject | Carcinoma, Non-Small-Cell Lung | |
dc.subject | Lung Neoplasms | |
dc.subject | Formaldehyde | |
dc.subject | DNA, Neoplasm | |
dc.subject | Paraffin Embedding | |
dc.subject | Tissue Fixation | |
dc.subject | Temperature | |
dc.subject | Gene Frequency | |
dc.subject | Mutation | |
dc.subject | Proto-Oncogene Proteins p21(ras) | |
dc.subject | High-Throughput Nucleotide Sequencing | |
dc.subject | Genotyping Techniques | |
dc.subject | Multiplex Polymerase Chain Reaction | |
dc.title | Efficient Genotyping of KRAS Mutant Non-Small Cell Lung Cancer Using a Multiplexed Droplet Digital PCR Approach. | |
dc.type | Journal Article | |
dcterms.dateAccepted | 2015-09-09 | |
rioxxterms.versionofrecord | 10.1371/journal.pone.0139074 | |
rioxxterms.licenseref.uri | https://creativecommons.org/licenses/by/4.0 | |
rioxxterms.licenseref.startdate | 2015-01 | |
rioxxterms.type | Journal Article/Review | |
dc.relation.isPartOf | PloS one | |
pubs.issue | 9 | |
pubs.notes | Not 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-status | Published | |
pubs.volume | 10 | |
pubs.embargo.terms | Not known | |
icr.researchteam | Molecular Oncology | |
icr.researchteam | Thoracic Oncology | |
icr.researchteam | Treatment of thoracic tumours | |
icr.researchteam | Lung Cancer Group | |
dc.contributor.icrauthor | Pender, Alexandra | |
dc.contributor.icrauthor | Garcia-Murillas, Isaac | |
dc.contributor.icrauthor | Cutts, Rosalind | |
dc.contributor.icrauthor | Turner, Nicholas | |