Epidermal Growth Factor Receptor (EGFR) Amplification May Lead to Invalid Cobas EGFR Mutation Test v2 Results
Abstract
:1. Introduction
2. Materials and Methods
2.1. Case Selection
2.2. Mutation Analysis
2.3. Epidermal Growth Factor Receptor (EGFR) Fluorescence In Situ Hybridization (FISH)
3. Results
3.1. The Invalid Rate Is 0.99% Among 4148 Cobas Epidermal Growth Factor Receptor (EGFR) Tests Conducted at the National Taiwan University Hospital (NTUH) from 2019 to 2022
3.2. Epidermal Growth Factor Receptor (EGFR) Amplification Resulted in a Low Cycle Threshold (Ct) Value for the Internal Control in the Four Invalid Cases, Leading to Invalid Results in the Cobas EGFR Test v2
3.3. All Four Cases with a Low Internal Control (IC) Cycle Threshold (Ct) Value Are Valid in the Second Round of Cobas Epidermal Growth Factor Receptor (EGFR) Testing Using Diluted Deoxyribonucleic Acid (DNA) Specimens
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Abbreviations
NSCLC | Non-small cell lung cancer |
EGFR | Epidermal Growth Factor Receptor |
IC | Internal Control |
NGS | Next-generation sequencing |
FISH | Fluorescence In Situ Hybridization |
References
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Year | Cobas Test “Valid” | Cobas Test “Invalid” | Prevalence |
---|---|---|---|
2019 | 718 | 14 | 1.91% |
2020 | 1184 | 10 | 0.84% |
2021 | 1373 | 6 | 0.44% |
2022 | 832 | 11 | 1.30% |
Total | 4107 | 41 | 0.99% |
Invalid Code | Description | Case: Total (Each Code) |
---|---|---|
R812/R832/R852 | Internal Control could not be detected | 11 (8/1/2) |
R813/R834/R853 | Internal Control out of range (High Ct) | 26 (24/1/1) |
R814 | Internal Control out of range (Low Ct) | 4 |
Total | 41 |
Age (Years) | Sex | Smoking | Tumor Site | Tumor Size (cm) | TNM (AJCC 8th) | |
---|---|---|---|---|---|---|
Case 1 | 59 | Male | Smoker | RUL | 7.5 | cT4N2M0 (stage IIIB) |
Case 2 | 72 | Male | Smoker | LUL | 4.4 | cT4N3M1c (stage IVB) |
Case 3 | 72 | Male | Non-smoker | RLL | 2.8 | cT4N3M1c (stage IVB) |
Case 4 | 54 | Male | Smoker | LUL | 10 | cT4N3M1a (stage IVA) |
Pathology | Tumor% | First Cobas Test | Sanger | Idylla | EGFR FISH | Second Cobas Test After Dilution of DNA Samples | NGS | |||
---|---|---|---|---|---|---|---|---|---|---|
SNV and InDels | EGFR CNV | MS | ||||||||
Case 1 | ADC (acinar, high-grade) | 30% | R814 | L858R | L858R (IC Ct 16.2) | Amp (large gene clusters) | L858R + Ex20ins (Ct: 31.16) | EGFR (p.L858R) NRAS (p.D176Y) TP53 (p.D281E) | Amp (27.33) | MS-Stable |
Case 2 | ADC (micropapillary) | 15% | R814 | WT | WT (IC Ct 17.9) | Amp (>15/cell) | Ex20ins (Ct: 31.01) | N/A | N/A | N/A |
Case 3 | ADC (micropapillary) | 30% | R814 | Ex19Del | Ex19Del (IC Ct 16.2) | Amp (large gene clusters) | Ex19Del + Ex20ins (Ct: 31.6) | EGFR ex19del (p.E746_ A750del) | Amp (16.75) | MS-Stable |
Case 4 | ADC (solid) | 25% | R814 | WT | T790M (IC Ct 14.8) | Amp (large gene clusters) | Ex20ins (Ct: 31.65) | TP53 (p.P153fs*28) | Amp (28.09) | MS-Stable |
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Hsieh, M.-S.; Hung, T.-C.; Huang, H.-N.; Lu, C.-W.; Hu, H.-W.; Lai, J.-Y.; Lee, W.-Y.; Chen, J.-S. Epidermal Growth Factor Receptor (EGFR) Amplification May Lead to Invalid Cobas EGFR Mutation Test v2 Results. Diagnostics 2025, 15, 948. https://doi.org/10.3390/diagnostics15080948
Hsieh M-S, Hung T-C, Huang H-N, Lu C-W, Hu H-W, Lai J-Y, Lee W-Y, Chen J-S. Epidermal Growth Factor Receptor (EGFR) Amplification May Lead to Invalid Cobas EGFR Mutation Test v2 Results. Diagnostics. 2025; 15(8):948. https://doi.org/10.3390/diagnostics15080948
Chicago/Turabian StyleHsieh, Min-Shu, Tze-Chun Hung, Hsien-Neng Huang, Chao-Wen Lu, Hsiang-Wei Hu, Jin-Yao Lai, Wen-Yao Lee, and Jin-Shing Chen. 2025. "Epidermal Growth Factor Receptor (EGFR) Amplification May Lead to Invalid Cobas EGFR Mutation Test v2 Results" Diagnostics 15, no. 8: 948. https://doi.org/10.3390/diagnostics15080948
APA StyleHsieh, M.-S., Hung, T.-C., Huang, H.-N., Lu, C.-W., Hu, H.-W., Lai, J.-Y., Lee, W.-Y., & Chen, J.-S. (2025). Epidermal Growth Factor Receptor (EGFR) Amplification May Lead to Invalid Cobas EGFR Mutation Test v2 Results. Diagnostics, 15(8), 948. https://doi.org/10.3390/diagnostics15080948