EGFR Mutation Analysis Kit

Available Products

Product Name/Description
No. of Reactions*
Product Code
EGFR Mutation Analysis Kit for Real-Time PCR
52
EGFR-RT52
EGFR Mutation Analysis Extension Kit †
52
EGFRX-RT52

* Includes all controls.

† For detection of E709, L858M, and L861R mutations.  Available with purchase of EGFR-RT52.

EGFR Mutations and Cancer

Epidermal growth factor receptor (EGFR) is a membrane protein that plays a central role in transmitting signals that promote cell growth and proliferation. Its tyrosine kinase (TK) domain activates several downstream effectors that lead to activation of the Ras-Raf-MAPK pathway. Overexpression and oncogenic mutations that constitutively activate the TK domain of EGFR have been found in various solid tumors. Additionally, excessive activation of EGFR has been shown to be associated with advanced stages of cancer and a poor prognosis.

Testing Procedure and Analysis

EntroGen’s EGFR mutation analysis kit is a real-time polymerase chain reaction (PCR)-based assay that uses mutant-specific probes to identify the presence of EGFR mutations. The testing procedure involves three (3) simple steps, which can be completed in approximately 2 hours from DNA to test results:

  • Isolation of DNA from tumor biopsies, paraffin-embedded sections (FFPE), fresh frozen tumors, or tumor cell lines.
  • PCR setup with reagents included in the kit.
  • Amplification (8 reactions per sample) and detection using a real-time PCR instrument.

Specifications

EntroGen’s EGFR mutation analysis kit detects the following EGFR mutations:

Exon 18

  • G719A – 2156G>C
  • G719S – 2155G>A
  • G719C – 2155G>T
  • G719D – 2156G>A

Exon 20

  • T790M – 2369C>T
  • S768I – 2303G>T
  • 2307-2308 ins GCCAGCGTG
  • 2319-2320 ins CAC
  • 2310-2311 ins GGT

Exon 19

  • 2235-2249 del 15
  • 2235-2252>AAT del 18
  • 2236-2253 del 18
  • 2237-2251 del 15
  • 2237-2254 del 18
  • 2237-2255>T del 19
  • 2236-2250 del 15
  • 2238-2255 del 18
  • 2238-2248>GC del 11
  • 2238-2252>GCA del 15
  • 2233-2247 del 15
  • 2234-2248 del 15
  • 2235-2246 del 12
  • 2235-2248>AATTC
  • 2235-2251>AATTC
  • 2235-2252>AAT
  • 2235-2255>AAT
  • 2236-2248>AGAC
  • 2236-2248>CAAC
  • 2236-2256 del 21
  • 2237-2252>T
  • 2239-2247 del 9
  • 2239-2256 del 18
  • 2239-2248>C del 10
  • 2239-2258>CA del 20
  • 2240-2251 del 12
  • 2240-2257 del 18
  • 2240-2254 del 15
  • 2239-2251>C del 13
  • 2237-2253>TC
  • 2237-2253>TTCCT
  • 2237-2253>TTGCT
  • 2237-2256>TC
  • 2237-2256>TT
  • 2237-2257>TCT
  • 2238-2252 del 15
  • 2239-2252>CA
  • 2239-2253 del 15
  • 2239-2256>CAA
  • 2239-2257>T
  • 2239-2262 del 24
  • 2246-2260 del 15
  • 2248-2273>CC
  • 2252-2275 del 24
  • 2252-2276>A
  • 2252-2277>AT
  • 2253-2276 del 24
  • 2254-2277 del 24

Exon 21

  • L858R – 2573T>G
  • L861Q – 2582T>A

EntroGen’s EGFR mutation analysis extension kit detects the following EGFR mutations:

Exon 18

  • E709
  • E709A
  • E709D (c.2127 A>C)
  • E709D (c.2127 A>T)
  • E709G
  • E709K
  • E709Q
  • E709V

Exon 20

  • L858M
  • L861R

Limitations

EntroGen’s EGFR mutation analysis kit requires 40-80 nanograms of genomic DNA (5-10 nanograms per reaction) and can detect a mutation load of less than 1%. This sensitivity greatly depends on the extent of fragmentation and quality of the isolated DNA.

Equipment and Materials

EntroGen’s EGFR mutation analysis kit requires a real-time PCR instrument capable of detecting FAM and VIC fluorescent probes.

This test includes reagents required for the PCR amplification/detection, as well as validated reaction controls. Columns and reagents for DNA isolation are not included.

Bibliography of studies using this product

Powrózek T, Krawczyk P, Pankowski J, Reszka K, Jakubiak M, Obrochta A, Wojas-Krawczyk K, Buczkowski J, Milanowski J.  Cytology smears as diagnostic material for EGFR gene testing in non-small cell lung cancer.  Tumori. 2015 Nov 14;101(6):e151-3. Pubmed ID 26108238 Lewandowska MA, Czubak K, Klonowska K, Jozwicki W, Kowalewski J, Kozlowski P. The Use of a Two-Tiered Testing Strategy for the Simultaneous Detection of Small EGFR Mutations and EGFR Amplification in Lung Cancer. PLoS One. 2015 Feb 26;10(2):e0117983. Pubmed ID 25719557 Rossella De Cecio, Monica Cantile, Francesca Collina, Laura Marra, Clemente Santonastaso, Cono Scaffa, Gerardo Botti, Nunzia Simona Losito. Borderline Brenner tumor of the ovary: a case report with immunohistochemical and molecular study. J Ovarian Res. 2014 Oct 29;7:101. Pubmed ID 25358264