Lung cancer EGFR — Labs

 

The therascreen EGFR RGQ PCR Kit reliably detects mutations in the EGFR gene, which are found in non-small cell lung cancer. According to the drug labels for GILOTRIF (afatinib) and IRESSA (gefitinib), EGFR (epidermal growth factor receptor) status evaluation must be detected by an FDA-approved test to determine treatment. The therascreen EGFR test is approved for in vitro diagnostic use in selecting patients for treatment with GILOTRIF (afatinib) or IRESSA (gefitinib) based on an EGFR mutation detected (EGFR positive) test result.
Information for Labs

 

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The therascreen EGFR test:

  • FDA-approved test for use in selecting patients with NSCLC in whom either GILOTRIF or IRESSA are indicated
  • Clinically validated in 2 trials, which together enrolled >450 patients* with NSCLC
  • Detects clinically relevant EGFR exon 19 deletions and L858R mutation
  • Belongs to an easy and complete workflow with automated reporting

Meet your oncologists’ request for a short turn around time by running the FDA-approved therascreen EGFR test in your lab.

* LUX-Lung 3 trial (1) with GILOTRIF (n=345) and the IFUM study (2) with IRESSA (n=106)


References

  1. Sequist, L.V. et al. (2013) Phase III study of afatinib or cisplatin plus pemetrexed in patients with metastatic lung adenocarcinoma with EGFR mutations. J. Clin. Oncol. 31:3327–3334

  2. Douillard J.Y. et al. (2014) First-line gefitinib in Caucasian EGFR mutation-positive NSCLC patients: a phase-IV, open-label, single-arm study. Br. J. Cancer 110, 55–62

Assay principle
Applications
Performance characteristics
therascreen assays in development
Reimbursement
Back to topAssay principle

The therascreen EGFR test provides eight separate PCR amplification reaction mixes: seven mutation specific reactions in exons 18, 19, 20 and 21 of the EGFR gene, and a wild-type control in exon 2. The principle components of the kit are explained below.

Mutation reaction mixes

Each mutation-specific reaction mix uses a mutation-specific ARMS primer to selectively amplify the mutated DNA, and then uses a Scorpions primer to detect the mutation during a second amplification reaction (see ARMS Scorpions technology).

ARMS

Allele-specific amplification is achieved by ARMS, which exploits the ability of Taq DNA polymerase to distinguish between a match and a mismatch at the 3' end of a PCR primer. When the primer is fully matched, the amplification proceeds with full efficiency. When the 3' base is mismatched, only low-level background amplification occurs. Therefore, a mutated sequence is selectively amplified even in samples where the majority of the DNA does not carry the mutation.

Scorpions

Detection of amplification is performed using Scorpions. Scorpions are bi-functional molecules containing a PCR primer covalently linked to a probe. The probes incorporate the fluorophore carboxyfluorescein (FAM) and a quencher. The latter quenches the fluorescence of the fluorophore. When the probe binds to the ARMS amplicon during PCR, the fluorophore and quencher become separated, leading to a detectable increase in fluorescence.


Back to top Applications

Mutations in the EGFR oncogene are found in human cancers (1-2). The presence of these mutations correlated with response to certain tyrosine kinase inhibitor (TKI) cancer therapies in patients with non-small cell lung cancer (NSCLC) (3-8). Such mutations in the EGFR oncogene are present at a frequency in the general population of patients with NSCLC at a frequency of about 10% in patients from the USA, Europe or Australia and up to 30% in patients from Japan and Taiwan (1, 2 and 9).

The therascreen EGFR RGQ PCR Kit is a real-time qualitative PCR assay used on the Rotor-Gene Q MDx instrument. Using Scorpions (10) and ARMS (Allele Refractory Mutation System) technologies (11), the therascreen EGFR RGQ PCR Kit enables the detection of 21 mutations in exons 18, 19, 20 and 21 of the EGFR oncogene against a background of wild-type genomic DNA (Table 1). The therascreen EGFR test detects 21 mutations in the EGFR oncogene which represent circa 82% of mutations reported in the COSMIC database (12).

Table 1. List of mutations and COSMIC identities
Mutation Exon Base change Cosmic ID
Deletions

19

2238_2255del18 6220
2235_2249del15 6223
2236_2250del15 6225
2239_2253del15 6254
2239_2256del18 6255
2240_2254del15 12369
2240_2257del18 12370
2239_2248TTAAGAGAAG>C 12382
2239_2251>C 12383
2237_2255>T 12384
2239_2258>CA 12387
2238_2252>GCA 12419
2238_2248>GC 12422
2235_2252>AAT 13551
L858R 21 2573T>G 6224
T790M* 20 2369C>T 6240
L861Q* 21 2582T>A 6213
G719A* 18 2156G>C 6239
S768I* 20 2303G>T 6241
Insertions* 20 2319_2320insCAC 12377
2310_2311insGGT 12378
*Safety and efficacy of GILOTRIF and IRESSA have not been established for these mutations.

References
  1. Pao, W., Miller, V.A. (2005) Epidermal growth factor receptor mutations, small molecule kinase inhibitors, and non-small-cell lung cancer: current knowledge and future directions. J. Clin. Oncol. 23, 2556.
  2. Johnson, B.E., and Jaenne, P.A. (2005) Epidermal growth factor receptor mutations in patients with non-small cell lung cancer. Cancer Res. 65, 7525.
  3. Inoue, A., Suzuki, T., Fukuhara, T., Maemondo, M., Kimura, Y. (2006) Prospective Phase II study of gefitinib for chemotherapy-naive patients with advanced non-small cell lung cancer with epidermal growth factor receptor gene mutations. J. Clin. Oncol. 24, 3340.
  4. Asahina, H. et al. (2006) A Phase II study of gefitinib as a first-line therapy for advanced non-small cell lung cancers with epidermal growth factor receptor (EGFR) gene mutations. 42nd Ann Mtg of the American Society of Clinical Oncology (ASCO), Atlanta 2 6 June 2006. J. Clin. Oncol. 24 (18S)(Suppl), Abstr 13014.
  5. Paz-Ares, L. et al. (2006) A prospective phase II trial of erlotinib in advanced non-small cell lung cancer (NSCLC) patients (p) with mutations in the tyrosine kinase (TK) domain of the epidermal growth factor receptor (EGFR). 42nd Ann Mtg of the American Society of Clinical Oncology (ASCO), Atlanta 2 6 June 2006. J. Clin. Oncol. 24 (18S)(Suppl), Abstr 7020.
  6. Kobayashi, K. et al. (2008) First-line gefitinib for poor PS patients with EGFR mutations. 44th Ann Mtg of the American Society of Clinical Oncology (ASCO), Chicago 31 May 3 June 2008. J. Clin. Oncol. (15S); 26 (15s)(Suppl), Abstr 8070.
  7. Sequist, L.V. et al. (2008) First-line gefitinib in patients with advanced non-small cell lung cancer harbouring somatic EGFR mutations. J. Clin. Oncol. 15, 2442.
  8. Porta,  R. et al. (2008) Erlotinib customization based on epidermal growth factor receptor (EGFR) mutations in stage IV non-small-cell lung cancer (NSCLC) patients (p). J. Clin. Oncol. 26 (May 20 suppl), Abstr 8038.
  9. Jaene, P.A., and Johnson, B.E. (2006) Effect of epidermal growth factor receptor tyrosine kinase domain mutation on the outcome of patients with non-small cell lung cancer treated with epidermal growth factor receptor tyrosine kinase inhibitors. rd Cambridge Con on Novel Agents in the Treatment of Lung Cancer: Advances in EGFR-Targeted Agents, Cambridge, 23-24 Sep 2005. Clin. Cancer Res. 12 (Suppl.14), 4416.
  10. Whitcombe, D. et al. (1999) Detection of PCR products using self-probing amplicons and fluorescence. Nature Biotech. 17, 804.
  11. Thelwell, N. et al. (2000) Mode of Action and Application of Scorpion Primers to Mutation Detection. Nucleic Acids Research 28, 3752.
  12. Catalog of Somatic Mutations in Cancer (www.sanger.ac.uk/genetics/CGP/cosmic).
Back to top Performance characteristics

The therascreen EGFR test provides:

  • Sensitive and specific PCR kit – detects EGFR mutations recommended by CAP and NCCN
  • Detection of 21 mutations in the EGFR oncogene present in circa 82% of EGFR-positive patients
  • 15 mutations have safety and efficacy of GILOTRIF and IRESSA established
  • Turn-around-time — testing can be performed in <8 hours
  • Easy workflow and software for automated report generation

 

Performance characteristics
Item Characteristic
Tissue section 5 µm section
Tumor content required 2 sections
Mutation coverage 21 mutations
Time to result Testing can be performed in <8 hours
Control/result interpretation Automatic
Limit of Detection — FFPE samples 0.81 – 16.87% for mutations with established safety and efficacy

Back to top therascreen assays in development

The Rotor-Gene Q MDx offers one platform for all future QIAGEN companion diagnostic (CDx) assays:

  • Rotor-Gene Q MDx instrument was FDA-approved as part of the EGFR testing system
  • Rotor-Gene Q MDx is a key element in the QIAGEN products

 


QIAGEN FDA-approved tests on RGQ MDX include:
  • therascreen EGFR RGQ PCR Kit for NSCLC validated with GILOTRIF and IRESSA
  • therascreen KRAS RGQ PCR Kit for mCRC validated with ERBITUX and Vectibix

Future therascreen tests for potential submission to FDA include:

  • EGFR for rociletinib
  • AKT
  • JAK2 V617F 
  • BCR-ABL1
  • PI3K

therascreen EGFR RGQ PCR Kit — for in vitro diagnostic use 


Back to top Reimbursement

The therascreen EGFR test is:

  • FDA-approved EGFR test for use in selecting patients with NSCLC in whom either GILOTRIF or IRESSA are indicated
  • Clinically validated in 2 trials, which together enrolled >450 patients* with NSCLC 
  • Sensitive and detects clinically relevant EGFR exon 19 deletions and L858R mutation
  • Easy to use and provides automated reporting

Meet your oncologists´ request for a short turn around time by running the FDA-approved therascreen EGFR test in your lab.

Public and private payers endorse the value of EGFR testing with strong coverage policies for approved indications. 
 
therascreen EGFR testing: 2015 coding and payment reference
2015 CPT code CPT code description
81235 EGFR (epidermal growth factor receptor; eg., non-small cell lung cancer) gene analysis, common variants (eg, exon 10 LREA deletion, L858R, T790M, G719A, G719S, L861Q)







Related ICD 9 codes Description
162.0 Malignant neoplasm of trachea
162.2 Malignant neoplasm of main bronchus
162.3 Malignant neoplasm of upper lobe, bronchus or lung
162.4 Malignant neoplasm of middle lobe, bronchus or lung
162.5 Malignant neoplasm of lower lobe, bronchus or lung
162.8 Malignant neoplasm of other parts of bronchus or lung
162.9 Malignant neoplasm of bronchus and lung, unspecified
197.0 Secondary malignant neoplasm of lung
V10.11 Personal history of malignant neoplasm of bronchus and lung















* LUX-Lung 3 trial (1) with GILOTRIF (n=345) and the IFUM study (2) with IRESSA (n=106)
References

 

  1. Sequist, L.V. et al. (2013) Phase III study of afatinib or cisplatin plus pemetrexed in patients with metastatic lung adenocarcinoma with EGFR mutations. J. Clin. Oncol. 31:3327–3334

  2. Douillard J.Y. et al. (2014) First-line gefitinib in Caucasian EGFR mutation-positive NSCLC patients: a phase-IV, open-label, single-arm study. Br. J. Cancer 110, 55–62


Questions

For technical support, please call:

800-DNA-PREP (800.362.7737)

For medical questions, please email our medical team:

medical_information@qiagen.com


QIAGEN has a broad portfolio of companion diagnostic products in development.






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External Links
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Clinical practice guidelines in oncology for patients with metastatic colorectal cancer.

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