title: Multiplex detection of hotspot mutations by rolling circle-enabled universal microarrays.
abstract: Detection of somatic low abundance mutations in early cancer development requires a discriminatory, specific, and high-throughput methodology. In this study we report specific, discriminatory detection of low abundance mutations through a novel combination of rolling circle amplification (Nat Genet 1998; 19:225-232) and PCR ligation detection reaction on a universal oligonucleotide microarray (J Mol Biol 1999; 292:251-262). After mutation-specific multiplex ligation and hybridization of 17 pairs of probes to a generic microarray, the ligated probes were visualized. The multiplex mutation-specific ligation is possible only because rolling circle amplification permits quantification of previously undetectable hybridization events conducive to the detection of a single mutation from within a pool of over 100 wild-type alleles. This system is readily adaptable to high-throughput automation using a robot such as the Biomek platform.
authors.affiliation: Department of Pathology, Yale New Haven Hospital, Yale University, New Haven, Connecticut, USA.
authors.forename: D P
authors.initials: DP
authors.lastname: Ladner
authors.forename: J H
authors.initials: JH
authors.lastname: Leamon
authors.forename: S
authors.initials: S
authors.lastname: Hamann
authors.forename: G
authors.initials: G
authors.lastname: Tarafa
authors.forename: T
authors.initials: T
authors.lastname: Strugnell
authors.forename: D
authors.initials: D
authors.lastname: Dillon
authors.forename: P
authors.initials: P
authors.lastname: Lizardi
authors.forename: J
authors.initials: J
authors.lastname: Costa
mesh_terms: D001331:Automation; D004273:DNA, Neoplasm / Q000032:analysis; D006801:Humans; D009154:Mutation; D009369:Neoplasms / Q000235:genetics*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D014407:Tumor Cells, Cultured
chemical_list: D004273:DNA, Neoplasm