Publication:
Implementation of novel pyrosequencing assays to screen for common mutations of BRAF and KRAS in a cohort of sporadic colorectal cancers

dc.contributor.author Packham, Deborah en_US
dc.contributor.author Ward, Robyn L en_US
dc.contributor.author Lin, Vita Ap en_US
dc.contributor.author Hawkins, Nicholas J en_US
dc.contributor.author Hitchins, Megan P en_US
dc.date.accessioned 2021-11-25T13:29:53Z
dc.date.available 2021-11-25T13:29:53Z
dc.date.issued 2009 en_US
dc.description.abstract Abstract: Activating mutations of the BRAF and KRAS genes cause constitutive stimulation of an important cell-signaling pathway promoting tumorigenesis, and are increasingly recognized as determinants of response to targeted cancer therapies. The V600E mutation accounts for most of the BRAF mutations in cancer, and KRAS mutations are predominantly encoded by nucleotide substitutions within codons 12 and 13. We designed novel pyrosequencing assays for the detection of the common ‘‘hotspot’’ mutations in these genes, which demonstrated analytical sensitivities of Z10% in titrations of mutant cell AQ1 lines. The KRAS pyrosequencing assay has the ability to simultaneously identify all potential nucleotide changes within the mutation cluster at codons 12 and 13, with a sequence output in the sense direction to facilitate results interpretation. These assays were used to determine the mutation status in a prospective series of 1198 sporadic colorectal cancers. The BRAF V600E mutation was detected in 13.2% of the colorectal cancers. The frequency of KRAS mutations in our cohort was 32.4%, with G>A transitions at position 2 of codons 12 and 13 being most prevalent. Both assays proved highly sensitive and specific when applied to clinical specimens, and were applicable to both fresh-frozen and formalin-fixed paraffin-embedded archival tissues. These assays would serve as a suitable platform for large-scale mutation detection in cancer specimens where the facility for pyrosequencing is available. en_US
dc.identifier.issn 1052-9551 en_US
dc.identifier.uri http://hdl.handle.net/1959.4/44373
dc.language English
dc.language.iso EN en_US
dc.rights CC BY-NC-ND 3.0 en_US
dc.rights.uri https://creativecommons.org/licenses/by-nc-nd/3.0/au/ en_US
dc.source Legacy MARC en_US
dc.title Implementation of novel pyrosequencing assays to screen for common mutations of BRAF and KRAS in a cohort of sporadic colorectal cancers en_US
dc.type Journal Article en
dcterms.accessRights metadata only access
dspace.entity.type Publication en_US
unsw.accessRights.uri http://purl.org/coar/access_right/c_14cb
unsw.identifier.doiPublisher http://dx.doi.org/10.1097/PDM.0b013e318182af52 en_US
unsw.relation.faculty Medicine & Health
unsw.relation.ispartofissue 2 en_US
unsw.relation.ispartofjournal Diagnostic Molecular Pathology en_US
unsw.relation.ispartofpagefrompageto 62-71 en_US
unsw.relation.ispartofvolume 18 en_US
unsw.relation.originalPublicationAffiliation Packham, Deborah, Clinical School - St Vincent's Hospital, Faculty of Medicine, UNSW en_US
unsw.relation.originalPublicationAffiliation Ward, Robyn L, Clinical School - St Vincent's Hospital, Faculty of Medicine, UNSW en_US
unsw.relation.originalPublicationAffiliation Lin, Vita Ap, Clinical School - St Vincent's Hospital, Faculty of Medicine, UNSW en_US
unsw.relation.originalPublicationAffiliation Hawkins, Nicholas J, Medical Sciences, Faculty of Medicine, UNSW en_US
unsw.relation.originalPublicationAffiliation Hitchins, Megan P, Clinical School - Prince of Wales Hospital, Faculty of Medicine, UNSW en_US
unsw.relation.school Clinical School St Vincents Hospital *
unsw.relation.school School of Medical Sciences *
unsw.relation.school Clinical School Prince of Wales Hospital *
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