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    Multiplexed assays for detection of mutations in PIK3CA.

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    Authors
    Board, Ruth E
    Thelwell, Nicola J
    Ravetto, Paul F
    Little, Stephen
    Ranson, Malcolm R
    Dive, Caroline
    Hughes, Andrew
    Whitcombe, David
    Affiliation
    Discovery Medicine, AstraZeneca Pharmaceuticals, Macclesfield, UK. ruth.board@astrazeneca.com
    Issue Date
    2008-04
    
    Metadata
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    Abstract
    BACKGROUND: Mutations in the PIK3CA gene (phosphoinositide-3-kinase, catalytic, alpha polypeptide) have recently been described in a number of cancers, and their detection is currently limited because of the low sensitivity of conventional sequencing techniques. METHODS: We combined Amplification Refractory Mutation System (ARMS; AstraZeneca) allele-specific PCR and Scorpions (DxS) to develop assays for tumor-borne PIK3CA mutations and used real-time PCR to develop high-throughput multiplexed assays for the most commonly reported PIK3CA mutants (H1047L, H1047R, E542K, E545K). RESULTS: These assays were more sensitive than sequencing and could detect 5 copies of mutant DNA in proportions as low as 0.1% of the total DNA. We assayed DNA extracted from human tumors and detected PIK3CA mutation frequencies of 10.2% in colorectal cancer, 38.7% in breast cancer, 1.9% in lung cancer, and 2.9% in melanoma. In contrast, sequencing detected only 53% of the mutations detected by our assay. CONCLUSIONS: Multiplexed assays, which can easily be applied to clinical samples, have been developed for the detection of PIK3CA mutations.
    Citation
    Multiplexed assays for detection of mutations in PIK3CA. 2008, 54 (4):757-60 Clin. Chem.
    Journal
    Clinical Chemistry
    URI
    http://hdl.handle.net/10541/65741
    DOI
    10.1373/clinchem.2007.098376
    PubMed ID
    18375489
    Type
    Article
    Language
    en
    ISSN
    0009-9147
    ae974a485f413a2113503eed53cd6c53
    10.1373/clinchem.2007.098376
    Scopus Count
    Collections
    All Christie Publications
    Clinical and Experimental Pharmacology Group
    Medical Oncology

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