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SALSA MLPA KIT P315 EGFR
[http://www.geneclinics.org/profiles/hht/details.html]
EGFR is a rational target in solid tumors. Activation of the EGFR promotes processes that are responsible for tumor growth and progression, including proliferation and maturation, angiogenesis, invasion, metastasis, and inhibition of apoptosis. In addition, EGFR expression has been detected to varying degrees in a wide range of solid tumors. Although the prognostic significance of EGFR expression remains unclear, a retrospective review of EGFR studies reported that EGFR expression levels are highly predictive of clinical outcome for patients with head and neck, ovarian, cervical, bladder, and esophageal cancers. They are of moderate prognostic value for gastric, breast, endometrial, and colorectal tumors. The mechanisms for oncogenic conversion of EGFR in cancer include amplified copy number, structural rearrangements of the receptor, and activating mutations that have all been detected in various malignancies. EGFR mutations cluster in the kinase domain of EGFR (exons 18-21), and cause ligand-independent activation of the receptor, representing possible targets for therapeutical intervention. In this regard, somatic EGFR mutations as well as gene amplification in patients with non-small cell lung cancer (NSCLC) highly correlate with the clinical response to tyrosine kinase inhibitors. The most common truncation is the EGFR deletion variant III (EGFRvIII). It contains an in-frame deletion of exons 2-7 from the extracellular domain of EGFR. The EGFR gene comprises 28 exons, spanning about 188 kb of genomic DNA and is located on 7p11 (53.5Mb from p-telomere). Most individuals with lung cancer have point mutations in the EGFR gene, most of which will not be detected by the MLPA technique. This P315-A1 EGFR probemix contains probes for each of the 28 exons of EGFR. Furthermore, it contains 9 reference probes in different chromosomal regions. Please note that some of these reference probes might be located in a region that is frequently deleted / amplified in a certain tumor type. This SALSA MLPA kit is designed to detect copy number changes of one or more exons of the EGFR gene. Heterozygote deletions of probe recognition sequences will be apparent by a 35-50% reduced relative peak area of the amplification product of that probe. However, mutations/polymorphisms very close to the probe ligation site may also result in a reduced relative peak area. Apparent deletions of a single exon therefore always require confirmation by other methods. We have no information on what percentage of defects in this gene is caused by copy number changes of complete exons. Please note that most defects in this gene are expected to be small (point) mutations, most of which will not be detected by this MLPA test.
Full mix description (pdf)
IMPORTANT NOTICE: MLPA kits are sold by MRC-Holland for research purposes and to demonstrate the possibilities of the MLPA technique. This kit is not CE/FDA certified for use in diagnostic procedures. Salsa MLPA kits are supplied with all necessary buffers and enzymes. Purchase of the Salsa MLPA test kits includes a limited license to use these products for research purposes. The use of this MLPA kit requires a thermocycler with heated lid and sequence type electrophoresis equipment. Different fluorescent PCR primers are available. The MLPA technique has been first described in Nucleic Acid Research 30, e57 (2002)
References
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Last change in probemix content Current lot number
: lot 0208 (February 2008) : lot 0208
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