Multiplex detection of ten ESR1 mutations and AKT1 E17K in breast cancer using digital PCR
Introduction: ESR1 mutations are now established as a key mechanism of resistance to endocrine therapy in estrogen-receptor–positive breast cancer (ER+ breast cancer) and their sensitive and specific detection in plasma-cell free DNA (plasma-cfDNA) is crucial to monitor during patient treatment. I...
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Elsevier
2024-09-01
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| Series: | The Journal of Liquid Biopsy |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2950195424000195 |
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| author | Stavroula Smilkou Aliki Ntzifa Dimitra Stergiopoulou Vasilis Georgoulias Evi Lianidou |
| author_facet | Stavroula Smilkou Aliki Ntzifa Dimitra Stergiopoulou Vasilis Georgoulias Evi Lianidou |
| author_sort | Stavroula Smilkou |
| collection | DOAJ |
| description | Introduction: ESR1 mutations are now established as a key mechanism of resistance to endocrine therapy in estrogen-receptor–positive breast cancer (ER+ breast cancer) and their sensitive and specific detection in plasma-cell free DNA (plasma-cfDNA) is crucial to monitor during patient treatment. In the present proof-of-principle study, we evaluated the performance of a novel multiplex assay (12plex) for the detection of ten ESR1 mutations and AKT1 E17K in plasma-cfDNA based on Crystal Digital PCR® (Stilla Technologies, France). Materials & methods: We analyzed 35 plasma-cfDNA samples from ER+ breast cancer patients and 10 samples from healthy donors and further compared the results with our previously reported ESR1 NAPA assay for D538G, Y537S, Y537C and Y537 N ESR1 mutations. Results: Using this novel 12plex ESR1-AKT 6-color Crystal Digital PCR® assay we detected both AKT1 E17K and ESR1 D538G mutations in 5/35 (14.3%) plasma-cfDNA samples. ESR1 D538G was detected in 4/35 (11.4%) of these plasma-cfDNA samples using the ESR1 NAPA assay. Direct comparison between Crystal Digital PCR™ and the ESR1 NAPA assay revealed a high concordance (97.1%, k = 0.871, p < 0.001) for the detection of D538G mutation. Conclusion: The Stilla 12plex ESR1-AKT 6-color Crystal Digital PCR® assay is multiplex, highly sensitive and robust and can be used in liquid biopsy. |
| format | Article |
| id | doaj-art-fbdae7aae3094e1cae2d264c9f3de265 |
| institution | Kabale University |
| issn | 2950-1954 |
| language | English |
| publishDate | 2024-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | The Journal of Liquid Biopsy |
| spelling | doaj-art-fbdae7aae3094e1cae2d264c9f3de2652024-11-22T07:41:30ZengElsevierThe Journal of Liquid Biopsy2950-19542024-09-015100154Multiplex detection of ten ESR1 mutations and AKT1 E17K in breast cancer using digital PCRStavroula Smilkou0Aliki Ntzifa1Dimitra Stergiopoulou2Vasilis Georgoulias3Evi Lianidou4Analysis of Circulating Tumor Cells, Laboratory of Analytical Chemistry, Department of Chemistry, University of Athens, 15771, Athens, GreeceAnalysis of Circulating Tumor Cells, Laboratory of Analytical Chemistry, Department of Chemistry, University of Athens, 15771, Athens, GreeceAnalysis of Circulating Tumor Cells, Laboratory of Analytical Chemistry, Department of Chemistry, University of Athens, 15771, Athens, GreeceFirst Department of Medical Oncology, Metropolitan General Hospital, Athens, 15562, GreeceAnalysis of Circulating Tumor Cells, Laboratory of Analytical Chemistry, Department of Chemistry, University of Athens, 15771, Athens, Greece; Corresponding author. National and Kapodistrian University of Athens, 15771, Athens, Greece.Introduction: ESR1 mutations are now established as a key mechanism of resistance to endocrine therapy in estrogen-receptor–positive breast cancer (ER+ breast cancer) and their sensitive and specific detection in plasma-cell free DNA (plasma-cfDNA) is crucial to monitor during patient treatment. In the present proof-of-principle study, we evaluated the performance of a novel multiplex assay (12plex) for the detection of ten ESR1 mutations and AKT1 E17K in plasma-cfDNA based on Crystal Digital PCR® (Stilla Technologies, France). Materials & methods: We analyzed 35 plasma-cfDNA samples from ER+ breast cancer patients and 10 samples from healthy donors and further compared the results with our previously reported ESR1 NAPA assay for D538G, Y537S, Y537C and Y537 N ESR1 mutations. Results: Using this novel 12plex ESR1-AKT 6-color Crystal Digital PCR® assay we detected both AKT1 E17K and ESR1 D538G mutations in 5/35 (14.3%) plasma-cfDNA samples. ESR1 D538G was detected in 4/35 (11.4%) of these plasma-cfDNA samples using the ESR1 NAPA assay. Direct comparison between Crystal Digital PCR™ and the ESR1 NAPA assay revealed a high concordance (97.1%, k = 0.871, p < 0.001) for the detection of D538G mutation. Conclusion: The Stilla 12plex ESR1-AKT 6-color Crystal Digital PCR® assay is multiplex, highly sensitive and robust and can be used in liquid biopsy.http://www.sciencedirect.com/science/article/pii/S2950195424000195Estrogen receptorBreast cancerESR1 mutationsCrystal digital PCRLiquid biopsyPlasma-cfDNA |
| spellingShingle | Stavroula Smilkou Aliki Ntzifa Dimitra Stergiopoulou Vasilis Georgoulias Evi Lianidou Multiplex detection of ten ESR1 mutations and AKT1 E17K in breast cancer using digital PCR The Journal of Liquid Biopsy Estrogen receptor Breast cancer ESR1 mutations Crystal digital PCR Liquid biopsy Plasma-cfDNA |
| title | Multiplex detection of ten ESR1 mutations and AKT1 E17K in breast cancer using digital PCR |
| title_full | Multiplex detection of ten ESR1 mutations and AKT1 E17K in breast cancer using digital PCR |
| title_fullStr | Multiplex detection of ten ESR1 mutations and AKT1 E17K in breast cancer using digital PCR |
| title_full_unstemmed | Multiplex detection of ten ESR1 mutations and AKT1 E17K in breast cancer using digital PCR |
| title_short | Multiplex detection of ten ESR1 mutations and AKT1 E17K in breast cancer using digital PCR |
| title_sort | multiplex detection of ten esr1 mutations and akt1 e17k in breast cancer using digital pcr |
| topic | Estrogen receptor Breast cancer ESR1 mutations Crystal digital PCR Liquid biopsy Plasma-cfDNA |
| url | http://www.sciencedirect.com/science/article/pii/S2950195424000195 |
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