PIK3CA Mutation Detection with Core Reagent A

Phosphoinositide-3-kinase (PI3K) plays an important role in the EGFR signaling pathway. The gene, PIK3CA, encoding the catalytic subunit of PI3K, has been found to be frequently mutated in human cancers including  colon cancer, gliomas, gastric cancer, breast cancer, endometrial cancer, and lung cancer1,2PIK3CA mutations in cancer cells are associated with early recurrence, poor prognosis and drug resistance. Analysis of PIK3CA mutations has been shown to be valuable in optimizing the anti-EGFR therapies for different types of cancers like lung cancer and metastatic colorectal cancers3-5.

Based on Shifted Termination Assay (STA) technology, MutectorTM reagents contain special chemistry and a modified enzyme mixture to enrich the mutation signal 20-fold and detect as low as 1% mutation in wild type background. MutectorTM reagents have been widely used to detect BRAF, KRAS and other mutations such as PIK3CA in clinical samples including FFPE samples (see STA/Mutector-related Publications).

The difference:

  • Single tube detects mutations in two exons –   Simultaneously detect 5 mutations (see below), detects all possible mutations in target codon
  • Use less clinical samples –  works for small samples such as fine needle aspiration (FNA)
  • Detects low level (1-5%) mutations
  • Accurate –  sequencing-like accuracy, the mutation is double confirmed by peak color and fragment size

Assay Information

Assay Format           One tube test   
Target Mutations      Exon 9: E542K,  F545G,  E545K
                                Exon20: H1047L, H1047R

Assay Platform         Capillary sequencer ABI 3100, 3700, 3130, 3500
Assay Sensitivity      Detects 1-2% PIK3CA mutations
Sample required       20-160 ng DNA 

Assay workflow

Sensitivity
Use 20 - 160ng DNA per reaction. These reagents detect 1-2% mutation(s) of total DNA and detect some mutations that are missed by sequencing 


Order Information

Cat. No. Product NameTarget MutationsSizeDocuments
Cor-ASTA Core Reagents A Universal Reagents for Point Mutations32 rxnProtocol

The gene specific primers are not included in the kit.    For research use only. Not for use in diagnostic procedures.

 

Core:  Cor-A

DNA Primer:  DP08

Data:

The MutectorTM  assay can simultaneously detect 5 PIK3CA mutations in the same tube:

Related Products

References

  1. Samuels Y et al. (2004). High frequency of mutations of the PIK3CA gene in human cancers. Science. 304:554
  2. Karakas B et al. (2006). Mutation of the PIK3CA oncogene in human cancers. Br J Cancer. 94:455
  3. Ligresti G et al. (2009). PIK3CA mutations in human solid tumors: role in sensitivity to various therapeutic approaches. Cell Cycle. 8:1352
  4. De Roock W et al. (2011). KRAS, BRAF, PIK3CA, and PTEN mutations: implications for targeted therapies in metastatic colorectal cancer. Lancet Oncol 12:594.
  5. Sequist LV et al. (2011). Genotypic and histological evolution of lung cancers acquiring resistance to EGFR inhibitors. Sci Transl Med. 3:75ra26