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EGFR Mutation Analysis PCR

EGFR, Lung cancer mutation analysis, non-small cell lung cancer

Test Codes

EPIC: LAB1231622, Beaker: EGFR

Department

Molecular Pathology

Specimen Collection Criteria

Collect: Paraffin-embedded tissue. A paraffin block must be submitted. (Slides or paraffin shavings are not acceptable.) Submit 10% formalin-fixed, paraffin-embedded block with corresponding H&E slide. Tissue should be well fixed and well processed. Average tissue size 5.0 mm2. Specimens requiring decalcification (e.g., bone biopsies) should be treated with Mol decal agent (EDTA) only. 

• The specimen must be accompanied by a completed requisition and must contain the patient name, date of birth, collection date, ordering physician, and source of specimen.

Physician Office/Draw Specimen Preparation

Maintain paraffin-embedded tissue at room temperature (20-25°C or 68-77°F) until transport.

Preparation for Courier Transport

Transport: Paraffin-embedded tissue, at room temperature (20-25°C or 68-77°F).

Rejection Criteria

• Tissue treated with decalcifying agents other than Mol Decal (EDTA).
• Fixatives other than 10% neutral buffered formalin (e.g. alcohol, zinc formalin). 
• Improper labeling or inadequate information.
• Less than 10 percent tumor cellularity, at discretion of medical director.
• Poor quality and/or quantity of extracted genomic DNA.

Testing will be cancelled on specimens meeting the above criteria with client notification.

Inpatient Specimen Preparation

Specimens at Royal Oak may be sent to the Surgical Pathology tube station, #201. In-house specimens are also picked up by a Surgical Pathology assistant every hour on the hour. Tissue blocks for in-house specimens are typically available in the department of Anatomic Pathology. Tissue blocks from outside institutions should be obtained via normal protocols for outside surgical specimens.

In-Lab Processing

Unstained sections of 5-µm thickness are cut from selected tissue blocks. The number of sections cut and the need for macro-dissection are determined by the pathologist, based upon the amount of available tissue and the tumor cellularity.

Maintain specimens at room temperature (20-25°C or 68-77°F) until testing.

Storage

Specimen Stability for Testing:

Room Temperature (20-25°C or 68-77°F); Indefinitely
Refrigerated (2-8°C or 36-46°F): Unacceptable
Frozen (-20°C/-4°F or below): Unacceptable
 
Specimen Storage, Tissue (post testing):

Room Temperature (20-25°C or 68-77°F): 7 days

Specimen Storage, DNA (post testing):

Frozen (-25 to -15°C): Indefinitely

Laboratory

Royal Oak Molecular Pathology Laboratory

Performed

Once per week.
Results available within 10 business days.

Reference Range

No EGFR mutation detected.

Test Methodology

Polymerase Chain Reaction (PCR) amplification of target sequences with real time fluorescent detection. 

This test detects EGFR mutations with a sensitivity of 1% in a background of wild type alleles; specific exceptions being: Exon 18 codon 719 (G719S, G719C, G719D and G719) and Exon 18 codon 709 (E709A, E709D, E709G, E709K, E709Q and E709V) which are detected with a sensitivity of 5% in a background of wild-type alleles. Mutation detection sensitivity corresponds to 2% and 10% tumor cells carrying a heterozygous mutation, respectively. This test is validated for use with formalin-fixed paraffin embedded tissue. The test performance has been established with surgical and cytology specimens containing pulmonary adenocarcinoma.

This assay detects the most prevalent somatic mutations in exons 18, 19, 20, and 21.

Nucleotide change	Amino acid change	Cosmic ID
Exon 18
c.2155G>A	p.G719S	6252
c.2155G>T	p.G719C	6253
c.2156G>A	p.G719D	18425
c.2156G>C	p.G719A	6239
Exon 19
c.2233_2247del15	p.K745_E749delKELRE	26038
c.2234_2248del15	p.K745_A750>T	1190791
c.2235_2246del12	p.E746_E749delELRE	28517
c.2235_2248>AATTC	p.E746_A750>IP	13550
c.2235_2249del15	p.E746_A750delELREA	6223
c.2235_2251>AATTC	p.E746_T751>IP	13552
c.2235_2252>AAT	p.E746_T751>I	13551
c.2235_2255>AAT	p.E746_S752>I	12385
c.2236_2248>AGAC	p.E746_A750>RP	12413
c.2236_2248>CAAC	p.E746_A750>QP	13557
c.2236_2250del15	p.E746_A750delELREA	6225
c.2236_2253del18	p.E746_T751delELREAT	12728
c.2236_2256del21	p.E746_S752delELREATS	133189
c.2237_2251del15	p.E746_T751>A	12678
c.2237_2251>TTC	p.E746_T751>VP	18421
c.2237_2252>T	p.E746_T751>V	12386
c.2237_2253>TC	p.E746_T751>V	133193
c.2237_2253>TTCCT	p.E746_T751>VP	52935
c.2237_2253>TTGCT	p.E746_T751>VA	12416
c.2237_2254del18	p.E746_S752>A	12367
c.2237_2255>T	p.E746_S752>V	12384
c.2237_2256>TC	p.E746_S752>V	18426
c.2237_2256>TT	p.E746_S752>V	133194
c.2237_2257>TCT	p.E746_P753>VS	18427
c.2238_2248>GC	p.L747_A750delinsP	12422
c.2238_2252>GCA	p.L747_T751delinsQ	12419
c.2240_2252del	p.L747_T751delLREAT	23571   to 12369
c.2238_2255del18	p.E746_S752>D	6220
c.2239_2247delTTAAGAGAA	p.L747_E749delLRE	6218
c.2239_2248TTAAGAGAAG>C	p.L747_A750>P	12382
c.2239_2251delinsC	p.L747_T751delinsP	12383
c.2239_2252delinsCA	p.L747_T751delinsQ	12420
c.2239_2252delinsCAA	p.L747_S752delinsQ	12403
c.2239_2253del15	p.L747_T751delLREAT	6254
c.2239_2256>CAA	p.L747_S752>Q	12403
c.2239_2256del18	p.L747_S752delLREATS	6255
c.2239_2257>T	p.L747_P753>S	133197
c.2239_2258>CA	p.L747_P753>Q	12387
c.2239_2262del24	p.L747_K754delLREATSPK	24970
c.2240_2251del12	p.L747_T751>S	6210
c.2240_2254del15	p.L747_T751delLREAT	12369
c.2240_2257del18	p.L747_P753>S	12370
c.2246_2260del15	p.A750_K754del	26718
c.2248_2273>CC	p.A750_E758>P	26440
c.2252_2275del24	p.T751_E758delTSPKANKE	133207
c.2252_2276>A	p.T751_I759>N	96856
c.2252_2277>AT	p.T751_I759>N	24270
c.2253_2276del24	p.S752_I759delSPKANKEI	13556
c.2254_2277del24	p.S752_I759delSPKANKEI	6256
Exon 20
Nucleotide change	Amino acid change	Cosmic ID
c.2369C>T	p.T790M	6240
c.2303G>T	p.S768I	6241
c.2307_2308insGCCAGCGTG	p.V769_D770insASV	12376
c.2310_2311insGGT	p.D770_N771insG	12378
c.2319_2320insCAC	p.H773_V774insH	12377
Exon 21
c.2573T>G	p.L858R	6224
c.2582T>A	p.L861Q	6213
Exon 18
Nucleotide change	Amino acid change	Cosmic ID
c.2126 A>C	E709A	13427
c.2127 A>C	E709D
c.2127 A>T	E709D
c.2126 G>C	E709G	13009
c.2125 G>A	E709K	12988
c.2125 G>C	E709Q	116882
c.2126 A>C	E709V	12371
Exon 21
Nucleotide change	Amino acid change	Cosmic ID
c.2572 C>A	p.L858M	12366
c.2582T>G	p.L861R	12374

Interpretation

--- POSITIVE; EGFR MUTATION(S) DETECTED.
--- NEGATIVE; NO EGFR MUTATION DETECTED.

Clinical Utility

The EGFR Mutation Analysis Assay is intended to assist clinicians in the selection of appropriate systemic therapy for advanced-stage non-small cell lung cancer patients by identifying mutations of EGFR exons 18, 19, 20, and 21 that confer either sensitivity or resistance to select tyrosine kinase inhibitors.

EGFR mutations typically occur in adenocarcinoma and rarely in squamous cell carcinoma. For carcinoma subtypes other than adenocarcinoma (i.e. squamous cell carcinoma), the utility of EGFR inhibitors in EGFR-mutated tumors is not well established.  Due to the large number of available biomarkers for this cancer, EGFR mutation is usually performed as part of an NGS panel in current practice. However, the Corewell Health East William Beaumont University Hospital institutional reflex policy calls for reflex EGFR testing in all resection specimens of non-small cell lung cancer. Additionally, isolated EGFR testing may also be performed upon clinician request in certain circumstances.

Per Current NCCN guidelines, EGFR testing should be performed in adenocarcinoma (and considered for squamous cell carcinoma) at the time of initial diagnosis for patients with stage IV disease and should be performed when adjuvant TKI therapy is a consideration for stage IB-IIIB disease.

CPT Codes

81235

Contacts

Last Updated

5/13/2025