--- Query Results ---

Program:Genetics and Genomics
Test Name:EGFR Amplification by FISH
Performing Site: University of Alberta Hospital
Performing Dept:Molecular Pathology
Availability:Test set-up on Tuesday.
TAT:2 weeks
Preferred Tube/Container:See Specimen Requirements
Specimen Requirements:For Paraffin-Embedded Tissue:
1. A formalin-fixed, paraffin-embedded tissue block is preferred.
2. Alternatively, one slide stained with Hematoxylin & Eosin as well as six 4-micron baked (2 hours at 60°C) unstained charged slides can be submitted.
3. Please provide a copy of the corresponding pathology report.
Requisition/Form:All requests MUST be submitted on a Molecular Pathology Requisition.
Please provide any pertinent clinical history on the requisition.
Indications:For the investigation of gliomas.
Method:Fluorescence In Situ Hybridization
Method Details:Slides are cut from paraffin block and are deparaffinized in xylene and dehydrated in ethanol. Specimen pretreatment is performed using SPoT-Light Tissue Pretreatment Kit (Life Technologies) and fluorescence in situ hybridization is performed using Vysis probes (Abbott Molecular Inc.).

Vysis EGFR / CEP 7 FISH Probes hybridize to chromosome 7p12 and chromosome 7p11.1q11.1 (alpha satellites), therefore detecting increased copy number and amplification of the EGFR gene, as well as chromosome 7 polysomy. In normal cells, there is an equal number of red and green signals (ratio EGFR to CEP7 <1.2), but in abnormal cells a ratio of EGFR to CEP7 >=1.2 and >=10% of cells showing more than 3 EGFR signals is indicative of EGFR amplification (1).

All appropriate negative and positive controls are used. An average of 100 interphase nuclei were examined for each probe set by two observers using an Olympus Provis Fluorescence microscope system on 1000X magnification.

(1) PTEN mutation, EGFR amplification and outcome in patients with anaplastic astrocytoma and GBM. JNCI J Natl Cancer Inst (2001) 93 (16): 1246-1256
Last Updated On:Wednesday, June 14, 2017
Date of Last Review:Jan 2 2018 12:00AM