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® CDx1,4,5 
Oncomine®Dx Target Test1,6
Guardant 360®1,7
 
  • Common EGFR mutations detected
  • Exon 19 deletions
  • L858R
  • T790M
  • Exon 19 deletions
  • L858R
  • T790M
  • Exon 19 deletions
  • L858R
  • T790M
  • Exon 19 deletions
  • L858R
  • Exon 19 deletions
  • L858R
  • T790M
  • Testing technology
  • qRT-PCR
  • qRT-PCR
  • NGS
  • NGS
  • NGS
  • Sample types
  • FFPE tissue
    Plasma
  • FFPE tissue
  • FFPE tissue
  • FFPE tissue
  • Plasma
  • Estimated turnaround time
  • Varies by testing laboratory
  • Varies by testing laboratory
  • <14 days
  • Varies by testing laboratory
  • 7 days

See the product label for more information on proper use of FDA-approved companion diagnostics.8

 

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® CDx1,4,5
  • 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
Guardant 360®1,7
  • Common EGFR mutations detected
  • Exon 19 deletions
  • L858R
  • T790M
  • Testing technology
  • NGS
  • Sample types
  • Plasma
  • Estimated turnaround time
  • 7 days

See the product label for more information on proper use of FDA-approved companion diagnostics.8

 

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

Assay technologies differ in turnaround time and sensitivity9

Assay technologies differ in turnaround time and sensitivity9

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® CDx1,4,10 Oncomine® Dx1,6 Guardant 360®1,7
 
  • Tumor content required
  • ≥10% tumor area
  • N/A
  • 20%-30% of tumor nuclei
  • ≥20%
  • N/A
  • 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
  • Two 10 mL tubes of blood
  • DNA required per test
  • 150 ng DNA (50 ng per reaction well)
  • N/A
  • 50-1000 ng DNA
  • 10 ng DNA
  • 5-30 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
  • 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
  • 2°C to 30°C, until printed expiration date
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 1,4,10
  • 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
Guardant 360®1,7
  • Tumor content required
  • N/A
  • Tissue section size
  • Two 10 mL tubes of blood
  • DNA required per test
  • 5-30 ng DNA
  • Mutation level
  • N/A
  • Reagents
  • 2°C to 30°C, until printed expiration date

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.11


Laboratory-developed tests can be used to detect EGFR mutations

A variety of LDTs are available

Listing in table below is not comprehensive

  GeneStrat®12,13 OncoBEAM™ 14-16
Target SelectorTM NGS Lung Panel17,18
 
  • Company
  • Biodesix
  • Sysmex Inostics
  • Biocept
  • Testing technology
  • PCR
  • PCR
  • NGS
  • Sample type
  • Blood
  • Blood 
  • Blood
  • Turnaround time
  • 3 days
  • 5-7 days
  • 7-10 days
GeneStrat®12,13
  • Company
  • Biodesix
  • Testing technology
  • PCR
  • Sample type
  • Blood
  • Turnaround time
  • 3 days
OncoBEAM™ 14-16
  • Company
  • Sysmex Inostics
  • Testing technology
  • PCR
  • Sample type
  • Blood
  • Turnaround time
  • 5-7 days
Target SelectorTM NGS Lung Panel17,18
  • Company
  • Biocept
  • Testing technology
  • NGS
  • Sample type
  • Blood
  • Turnaround time
  • 7-10 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 September 10, 2020. Accessed September 15, 2020. 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. Guardant 360® CDx [technical information]. Redwood City, CA: Guardant Health, Inc; 2020. 8. FDA. https://www.fda.gov/downloads/MedicalDevices/DeviceRegulationandGuidance/GuidanceDocuments/UCM262327.pdf. Published August 6, 2014. Accessed March 16, 2018. 9. Lindeman NI et al. Arch Pathol Lab Med. 2018.142(3):321-346. doi:10.5858/arpa.2017-0388-CP. 10. FoundationOne® CDx [specimen instructions]. Cambridge, MA: Foundation Medicine, Inc.; 2017. 11. Levy BP et al. Oncologist. 2015;20(10):1175-1181. 12. 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. 13. Biodesix. https://www.biodesix.com/products/lung-cancer/lung-reflex/genestrat. Accessed July 7, 2020. 14. GenomeWeb. https://www.genomeweb.com/pcrsample-prep/inostics-lab-gains-clia-licensure. Published May 30, 2013. Accessed March 16, 2018. 15. Sysmex Inostics. https://www.oncobeam.com/healthcare-providers/lung-cancer. Accessed July 7, 2020. 16. Sysmex Inostics. https://www.oncobeam.com/oncobeam-technology/technology-overview. Accessed July 7, 2020. 17. Biocept. https://biocept.com/about/. Accessed July 7, 2020. 18. Biocept. https://biocept.com/ngs-lung-panel. Accessed June 24, 2020. 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.