title: Sensitive quantification of somatic mutations using molecular inversion probes.
abstract: Somatic mutations in DNA can serve as cancer specific biomarkers and are increasingly being used to direct treatment. However, they can be difficult to detect in tissue biopsies because there is often only a minimal amount of sample and the mutations are often masked by the presence of wild type alleles from nontumor material in the sample. To facilitate the sensitive and specific analysis of DNA mutations in tissues, a multiplex assay capable of detecting nucleotide changes in less than 150 cells was developed. The assay extends the application of molecular inversion probes to enable sensitive discrimination and quantification of nucleotide mutations that are present in less than 0.1% of a cell population. The assay was characterized by detecting selected mutations in the KRAS gene, which has been implicated in up to 25% of all cancers. These mutations were detected in a single multiplex assay by incorporating the rapid flow cytometric readout of multiplexable DNA biosensors.
authors.affiliation: Centre for Biomarker Research and Development, Australian Institute for Bioengineering and Nanotechnology (AIBN), The University of Queensland, Brisbane, QLD, Australia.
authors.forename: Rena
authors.initials: R
authors.lastname: Hirani
authors.forename: Ashley R
authors.initials: AR
authors.lastname: Connolly
authors.forename: Lisa
authors.initials: L
authors.lastname: Putral
authors.forename: Alexander
authors.initials: A
authors.lastname: Dobrovic
authors.forename: Matt
authors.initials: M
authors.lastname: Trau
mesh_terms: D015374:Biosensing Techniques; D007446:Chromosome Inversion; D004252:DNA Mutational Analysis; D017931:DNA Primers / Q000737:chemistry; D004273:DNA, Neoplasm / Q000235:genetics*; D005434:Flow Cytometry; D006801:Humans; D015335:Molecular Probes; D009154:Mutation / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D016283:Proto-Oncogene Proteins p21(ras); D018631:ras Proteins / Q000235:genetics*
chemical_list: D017931:DNA Primers; D004273:DNA, Neoplasm; C117307:KRAS protein, human; D015335:Molecular Probes; D011518:Proto-Oncogene Proteins; D016283:Proto-Oncogene Proteins p21(ras); D018631:ras Proteins