AuthorsBoard, Ruth E
Thelwell, Nicola J
Ravetto, Paul F
Ranson, Malcolm R
AffiliationDiscovery Medicine, AstraZeneca Pharmaceuticals, Macclesfield, UK. firstname.lastname@example.org
MetadataShow full item record
AbstractBACKGROUND: 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.
CitationMultiplexed assays for detection of mutations in PIK3CA. 2008, 54 (4):757-60 Clin. Chem.
- Detection of PIK3CA gene mutations with HRM analysis and association with IGFBP-5 expression levels in breast cancer.
- Authors: Dirican E, Kaya Z, Gullu G, Peker I, Ozmen T, Gulluoglu BM, Kaya H, Ozer A, Akkiprik M
- Issue date: 2014
- Novel mutant-enriched sequencing identified high frequency of PIK3CA mutations in pharyngeal cancer.
- Authors: Qiu W, Tong GX, Manolidis S, Close LG, Assaad AM, Su GH
- Issue date: 2008 Mar 1
- Detection of PIK3CA Mutations in Plasma DNA of Colorectal Cancer Patients by an Ultra-Sensitive PNA-Mediated PCR.
- Authors: Zeng Q, Xie L, Zhou N, Liu M, Song X
- Issue date: 2017 Aug
- Molecular evaluation of PIK3CA gene mutation in breast cancer: determination of frequency, distribution pattern and its association with clinicopathological findings in Indian patients.
- Authors: Ahmad F, Badwe A, Verma G, Bhatia S, Das BR
- Issue date: 2016 Jul
- Highly sensitive detection of the PIK3CA (H1047R) mutation in colorectal cancer using a novel PCR-RFLP method.
- Authors: Li WM, Hu TT, Zhou LL, Feng YM, Wang YY, Fang J
- Issue date: 2016 Jul 12