EGFR Diagnostic Assays

Multiple FDA-approved assays are available

FDA-approved assays utilize different technologies

  cobas® EGFR Mutation
Test v21,2 
therascreen® EGFR RGQ
PCR Kit1,3 
FoundationOne CDx™ 1,4,5 
Oncomine™ Dx Target Test1,6
 
  • Common EGFR mutations detected
  • Exon 19 deletions
  • L858R
  • T790M
  • Exon 19 deletions
  • L858R
  • T790M
  • Exon 19 deletions
  • L858R
  • T790M
  • Exon 19 deletions
  • L858R
  • Testing technology
  • qRT-PCR
  • qRT-PCR
  • NGS
  • NGS
  • Sample types
  • FFPE tissue
    Plasma
  • FFPE tissue
  • FFPE tissue
  • FFPE tissue
  • Estimated turnaround time
  • Varies by testing laboratory
  • Varies by testing laboratory
  • <14 days
  • Varies by testing laboratory

See the product label for more information on proper use of FDA-approved companion diagnostics.7

 

cobas® EGFR Mutation Test v21,2 
  • Common EGFR mutations detected
  • Exon 19 deletions
  • L858R
  • T790M
  • Testing technology
  • qRT-PCR
  • Sample types
  • FFPE tissue
    Plasma
  • Estimated turnaround time
  • Varies by testing laboratory
therascreen® EGFR RGQ PCR Kit1,3 
  • Common EGFR mutations detected
  • Exon 19 deletions
  • L858R
  • T790M
  • Testing technology
  • qRT-PCR
  • Sample types
  • FFPE tissue
  • Estimated turnaround time
  • Varies by testing laboratory
FoundationOne CDxTM 1,4,5
  • Common EGFR mutations detected
  • Exon 19 deletions
  • L858R
  • T790M
  • Testing technology
  • NGS
  • Sample types
  • FFPE tissue
  • Estimated turnaround time
  • <14 days
Oncomine Dx Target Test1,6
  • Common EGFR mutations detected
  • Exon 19 deletions
  • L858R
  • Testing technology
  • NGS
  • Sample types
  • FFPE tissue
  • Estimated turnaround time
  • Varies by testing laboratory

See the product label for more information on proper use of FDA-approved companion diagnostics.7

 

PCR- and NGS-based assays have different benefits and drawbacks

Assay technologies differ in turnaround time and sensitivity8

Assay technologies differ in turnaround time and sensitivity8

qRT-PCR
NGS
 
  • Selectively amplifies a target DNA sequence and quantifies the amount of target DNA
  • Can sequence entire genomes or specific areas of interest
  • May have a shorter turnaround time
  • May have a longer turnaround time
  • May be less sensitive
  • May be more sensitive


Sample requirements vary by assay

Sample requirements for FDA-approved tests

  cobas® EGFR
Mutation Test v21,2
therascreen® EGFR
RGQ PCR Kit1,3
FoundationOne CDx™ 1,4,9 Oncomine™ Dx1,6
 
  • Tumor content required
  • ≥10% tumor area
  • N/A
  • 20%-30% of tumor nuclei
  • ≥20%
  • Tissue section size
  • 5-µm section
  • 2 slides/5 µm per slide
    (per extraction)
  • FFPE block + 1 H&E slide or
    10 slides/4-5 µm per slide
  • 2-9 slides/5 µm per slide
  • DNA required per test
  • 150 ng DNA (50 ng per reaction well)
  • N/A
  • 50-1000 ng DNA
  • 10 ng DNA
  • Mutation level
  • ≥5% in 50 ng per reaction well
  • Exon 19 del – variable
  • L858R – 5.94%
  • T790M – 17.5%
  • 1.8%-3.4%
    (dependent on EGFR mutation)
  • N/A
  • Reagents
  • Refrigerated 2°C-8°C, liquid ready to use; up to 90 days open vial stability
  • N/A
  • N/A
  • Target Test: −30°C to −10°C
  • Target DNA Control: −30°C to −10°C
  • Target RNA Control: −90°C to −60°C
  • No Template Control: 15°C
    to 30°C
cobas® EGFR Mutation Test v21,2 
  • Tumor content required
  • ≥10% tumor area
  • Tissue section size
  • 5-µm section
  • DNA required per test
  • 150 ng DNA (50 ng per reaction well)
  • Mutation level
  • ≥5% in 50 ng per reaction well
  • Reagents
  • Refrigerated 2°C-8°C, liquid ready to use; up to 90 days open vial stability
therascreen® EGFR RGQ PCR Kit1,3 
  • Tumor content required
  • N/A
  • Tissue section size
  • 2 slides/5 µm per slide
    (per extraction)
  • DNA required per test
  • N/A
  • Mutation level
  • Exon 19 del – variable
  • L858R – 5.94%
  • T790M – 17.5%
  • Reagents
  • N/A
FoundationOne CDx 1,4,5
  • Tumor content required
  • 20%-30% of tumor nuclei
  • Tissue section size
  • FFPE block + 1 H&E slide or 10 slides/4-5 µm per slide
  • DNA required per test
  • 50-1000 ng DNA
  • Mutation level
  • 1.8%-3.4% (dependent on EGFR mutation)
  • Reagents
  • N/A
Oncomine Dx1,6
  • Tumor content required
  • ≥20%
  • Tissue section size
  • 2-9 slides/5 µm per slide
  • DNA required per test
  • 10 ng DNA
  • Mutation level
  • N/A
  • Reagents
  • Target Test: −30°C to −10°C
  • Target DNA Control: −30°C to −10°C
  • Target RNA Control: −90°C to −60°C
  • No Template Control: 15°C to 30°C

Sample collection method may impact the ability to test for biomarkers. Consult with your multidisciplinary team to ensure collection methods are appropriate for the planned diagnostic tests.10


Laboratory-developed tests can be used to detect EGFR mutations

A variety of LDTs are available

  Guardant 360™ 11,12 GeneStrat®13,14 OncoBEAM™ 15,16
ExoDx Lung17 Biocept Liquid Biopsy18
 
  • Company
  • Guardant
  • Biodesix
  • Sysmex Inostics
  • Exosome Diagnostics
  • Biocept
  • Testing technology
  • NGS
  • PCR
  • PCR
  • PCR
  • PCR
  • Sample type
  • Blood
  • Blood
  • Tissue
    Blood 
  • Blood
  • Blood
  • Turnaround time
  • 14 days
  • 3 days
  • <10 days
  • <7 days
  • <7 days
Guardant 360 11,12
  • Company
  • Guardant
  • Testing technology
  • NGS
  • Sample type
  • Blood
  • Turnaround time
  • 14 days
GeneStrat® 13,14
  • Company
  • Biodesix
  • Testing technology
  • PCR
  • Sample type
  • Blood
  • Turnaround time
  • 3 days
OncoBEAM™ 15,16
  • Company
  • Sysmex Inostics
  • Testing technology
  • PCR
  • Sample type
  • Tissue
    Blood
  • Turnaround time
  • <10 days
ExoDx Lung17   
  • Company
  • Exosome Diagnostics
  • Testing technology
  • PCR
  • Sample type
  • Blood
  • Turnaround time
  • <7 days
Biocept Liquid Biopsy18   
  • Company
  • Biocept
  • Testing technology
  • PCR
  • Sample type
  • Blood
  • Turnaround time
  • <7 days

LDTs were developed and their performance characteristics determined by specific institutions in a manner consistent with the Clinical Laboratory Improvement Amendments of 1988 (CLIA) requirements.19,20,*

Sample requirements vary for each LDT. Always verify that your samples adhere to the individual test specifications.

 

*These tests are performed in laboratories that adhere to performance specifications established by the Clinical Laboratory Improvement Amendments of 1988 (CLIA) under the regulation of the Centers for Medicare & Medicaid Services. They are not FDA approved.

AMP, Association for Molecular Pathology; CAP, College of American Pathologists; EGFR, epidermal growth factor receptor; FDA, Food and Drug Administration; FFPE, formalin-fixed paraffin-embedded; H&E, hematoxylin and eosin; IASLC, International Association for the Study of Lung Cancer; LDT, laboratory-developed test; mNSCLC, metastatic non–small cell lung cancer; N/A, not available; NGS, next-generation sequencing; PCR, polymerase chain reaction; qRT-PCR, quantitative real-time PCR.

References: 1. Food and Drug Administration (FDA). http://www.fda.gov/MedicalDevices/ProductsandMedicalProcedures/InVitroDiagnostics/ucm301431.htm. Updated January 23, 2018. Accessed March 16, 2018. 2. cobas® EGFR Mutation Test v2 [package insert]. Branchburg, NJ: Roche Molecular Systems, Inc.; 2015. 3. therascreen® EGFR RGQ PCR Kit [instructions for use (handbook)]. Manchester, UK: QIAGEN; 2017. 4. FoundationOne CDx™ [technical specifications]. Cambridge, MA: Foundation Medicine, Inc.; 2017. 5. Foundation Medicine. https://www.foundationmedicine.com/genomic-testing/foundation-one-cdx. Accessed March 16, 2018. 6. Oncomine™ Dx Target Test Part I: sample preparation and quantification [User Guide]. Waltham, MA: Thermo Fisher Scientific Inc.; 2017. 7. FDA. https://www.fda.gov/downloads/MedicalDevices/DeviceRegulationandGuidance/GuidanceDocuments/UCM262327.pdf. Published August 6, 2014. Accessed March 16, 2018. 8. Lindeman NI et al. Arch Pathol Lab Med. 2018.142(3):321-346. doi:10.5858/arpa.2017-0388-CP. 9. FoundationOne CDx™ [specimen instructions]. Cambridge, MA: Foundation Medicine, Inc.; 2017. 10. Levy BP et al. Oncologist. 2015;20(10):1175-1181. 11. Guardant Health. http://www.guardanthealth.com/medical-professionals/#gene-panel. Accessed March 16, 2018. 12. Guardant Health. http://www.guardanthealth.com/guardant360/#how-it-works. Accessed March 16, 2018. 13. Centers for Medicare & Medicaid Services (CMS). https://www.biodesix.com/wp-content/uploads/2017/09/CLIA-Accreditation-cert-exp-6-20-2019-1.pdf. Accessed March 16, 2018. 14. Biodesix. https://www.biodesix.com/genestrat. Accessed March 16, 2018. 15. GenomeWeb. https://www.genomeweb.com/pcrsample-prep/inostics-lab-gains-clia-licensure. Published May 30, 2013. Accessed March 16, 2018. 16. Sysmex Inostics. https://www.sysmex-inostics.com/our-services/product-single-view/oncobeamTM-egfr-3805.html. Accessed March 16, 2018. 17. ExoDx® Lung(ALK) [fact sheet]. Waltham, MA: Exosome Diagnostics, Inc; 2016. http://www.exosomedx.com/sites/default/files/uploads/alk_t790m_051716_0.pdf. Accessed March 16, 2018. 18. Biocept. https://biocept.com/technology/lung-cancer-offering. Accessed March 16, 2018. 19. Fitzgibbons PL et al. Arch Pathol Lab Med. 2014;138(11):1432-1443. 20. FDA. https://www.fda.gov/medicaldevices/productsandmedicalprocedures/invitrodiagnostics/laboratorydevelopedtests/default.htm. Updated February 4, 2018. Accessed March 16, 2018. 21. Lindeman NI et al. Arch Pathol Lab Med. 2013;137(6):828-860.