A Highly Sensitive Next-Generation Sequencing-Based Genotyping Platform for EGFR Mutations in Plasma from Non-Small Cell Lung Cancer Patients
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Study Population
2.2. Sample Preparation
2.3. EGFR Mutation Detection Platforms
2.4. Sel-Cap Mutation Enrichment PCR
2.5. NGS Library Preparation
2.6. Monitoring EGFR T790M in Plasma for EGFR-TKI Treatment
2.7. Statistical Analyses
3. Results
3.1. Study Population
3.2. Sel-Cap Showed High Sensitivity for EGFR Mutations in Plasma
3.3. Timing of First T790M Detection in Plasma is Critical for PFS of First-Line EGFR-TKIs
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Ex19del | L858R | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Sel-Cap | NGS | Sel-Cap | NGS | ||||||||||
Mutant | Wild | Total | Mutant | Wild | Total | Mutant | Wild | Total | Mutant | Wild | Total | ||
PNAclamp | Mutant | 15 | 5 | 20 | 3 | 15 | 18 | 11 | 6 | 17 | 2 | 14 | 16 |
Wild | 2 | 39 | 41 | 1 | 42 | 43 | 0 | 44 | 44 | 0 | 45 | 45 | |
Total | 17 | 44 | 61 | 4 | 57 | 61 | 11 | 50 | 61 | 2 | 59 | 61 | |
Sensitivity | (95% CI) | 75% | (53–89%) | 17% | (6–39%) | 65% | (41–83%) | 13% | (3–36%) | ||||
Specificity | (95% CI) | 95% | (84–99%) | 98% | (88–100%) | 100% | (92–100%) | 100% | (92–100%) | ||||
Accuracy | (95% CI) | 89% | (78–94%) | 74% | (62–83%) | 90% | (80–95%) | 77% | (65–86%) | ||||
Kappa | (95% CI) | 0.73 | (0.54–0.92) | 0.19 | (−0.16–0.53) | 0.73 | (0.52–0.93) | 0.17 | (−0.21–0.55) |
Sel-Cap | Ex19del | L858R | T790M a | |||||||
---|---|---|---|---|---|---|---|---|---|---|
Mutant | Wild | Total | Mutant | Wild | Total | Mutant | Wild | Total | ||
PNAclamp | Mutant | 50 | 19 | 69 | 24 | 12 | 36 | 7 | 14 | 21 |
Wild | 1 | 115 | 116 | 3 | 146 | 149 | 1 | 63 | 64 | |
Total | 51 | 134 | 185 | 27 | 158 | 185 | 8 | 77 | 85 | |
Sensitivity | (95% CI) | 72% | (61–82%) | 67% | (50–80%) | 88% | (53–98%) | |||
Specificity | (95% CI) | 99% | (95–100%) | 98% | (94–99%) | 82% | (72–89%) | |||
Accuracy | (95% CI) | 89% | (84–93%) | 92% | (87–95%) | 82% | (73–89%) | |||
Kappa | (95% CI) | 0.76 | (0.65–0.86) | 0.71 | (0.57–0.85) | 0.40 | (0.13–0.68) |
No. | EGFR-TKI | Tumor | Plasma | |
---|---|---|---|---|
PNAclamp | NGS Cancer Panel | Sel-Cap | ||
1 | Erlotinib | Ex19del a | Ex19del | Ex19del, T790M d |
2 | Erlotinib | Ex19del a | Wild | T790M d |
3 | Erlotinib | L858R a | L858R, T790M d | Wild |
4 | Erlotinib | Ex19del a | Wild | T790M d |
5 | Erlotinib | Ex19del a | Ex19del d | Wild |
6 | Gefitinib | Ex19del, T790M a | Wild | Wild |
7 | Gefitinib | Ex19del a | Ex19del, T790M | Ex19del, T790M |
8 | Afatinib | Ex19del a | Wild | Wild |
9 | Afatinib | Ex19del a | Ex19del | Ex19del, T790M d |
10 | Gefitinib | L858R, T790M b | L858R, T790M d | L858R |
11 | Gefitinib | Ex19del, T790M b | Wild | Wild |
12 | Afatinib | Ex19del b | Ex19del | Ex19del, T790M d |
13 | Erlotinib | T790M c | Wild | Wild |
14 | Erlotinib | T790M c | Wild | Wild |
15 | Erlotinib | L858R, T790M c | L858R | L858R, T790M d |
16 | Erlotinib | Ex19del, T790M c | Ex19del | Ex19del |
17 | Erlotinib | Ex19del, T790M c | Wild | Ex19del, T790M d |
18 | Erlotinib | Ex19del, T790M c | Ex19del | Ex19del, T790M d |
19 | Gefitinib | Ex19del, T790M c | Wild | Ex19del, T790M d |
20 | Gefinitib | Ex19del, T790M c | Wild | Ex19del, T790M d |
21 | Gefinitib | L858R, T790M c | L858R, T790M | L858R, T790M |
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Shin, J.-Y.; Kim, J.-O.; Lee, M.-R.; Kim, S.R.; Beck, K.S.; Kang, J.H. A Highly Sensitive Next-Generation Sequencing-Based Genotyping Platform for EGFR Mutations in Plasma from Non-Small Cell Lung Cancer Patients. Cancers 2020, 12, 3579. https://doi.org/10.3390/cancers12123579
Shin J-Y, Kim J-O, Lee M-R, Kim SR, Beck KS, Kang JH. A Highly Sensitive Next-Generation Sequencing-Based Genotyping Platform for EGFR Mutations in Plasma from Non-Small Cell Lung Cancer Patients. Cancers. 2020; 12(12):3579. https://doi.org/10.3390/cancers12123579
Chicago/Turabian StyleShin, Jung-Young, Jeong-Oh Kim, Mi-Ran Lee, Seo Ree Kim, Kyongmin Sarah Beck, and Jin Hyoung Kang. 2020. "A Highly Sensitive Next-Generation Sequencing-Based Genotyping Platform for EGFR Mutations in Plasma from Non-Small Cell Lung Cancer Patients" Cancers 12, no. 12: 3579. https://doi.org/10.3390/cancers12123579