Norimitsu Kasahara1, Hirotsugu Kenmotsu2, Masakuni Serizawa3, Rina Umehara4, Akira Ono5, Yasushi Hisamatsu6, Kazushige Wakuda5, Shota Omori5, Kazuhisa Nakashima5, Tetsuhiko Taira7, Tateaki Naito5, Haruyasu Murakami5, Yasuhiro Koh8, Keita Mori9, Masahiro Endo10, Takashi Nakajima11, Masanobu Yamada12, Masatoshi Kusuhara4, Toshiaki Takahashi5. 1. Division of Thoracic Oncology, Shizuoka Cancer Center, Japan; Department of Medicine and Molecular Science, Gunma University Graduate School of Medicine, 3-39-15, Showa-machi, Maebashi, Gunma 371-8511, Japan. 2. Division of Thoracic Oncology, Shizuoka Cancer Center, Japan. Electronic address: h.kenmotsu@scchr.jp. 3. Drug Discovery and Development Division, Shizuoka Cancer Center Research Institute, Japan. Electronic address: m.serizawa@scchr.jp. 4. Drug Discovery and Development Division, Shizuoka Cancer Center Research Institute, Japan. 5. Division of Thoracic Oncology, Shizuoka Cancer Center, Japan. 6. Division of Thoracic Oncology, Shizuoka Cancer Center, Japan; Department of Medical Oncology and Hematology, Oita University Faculty of Medicine, 1-1 Idaigaoka, Hasamamachi, Yufu, Oita, 879-5593, Japan. 7. Division of Thoracic Oncology, Shizuoka Cancer Center, Japan; Division of Respiratory Medicine, Minami Kyushu National Hospital, 1882 Kida, Kajiki-chou, Aira, Kagoshima, 899-5293, Japan. 8. Third Department of Internal Medicine, Wakayama Medical University, 811-1 Kimiidera, Wakayama, Wakayama, 641-8509, Japan. 9. Clinical Trial Coordination Office, Shizuoka Cancer Center, Japan. 10. Division of Diagnostic Radiology, Shizuoka Cancer Center, Japan. 11. Division of Diagnostic Pathology, Shizuoka Cancer Center, 1007 Shimonagakubo, Nagaizumi-chou, Suntou-gun, Shizuoka, 411-8777, Japan. 12. Department of Medicine and Molecular Science, Gunma University Graduate School of Medicine, 3-39-15, Showa-machi, Maebashi, Gunma 371-8511, Japan.
Abstract
OBJECTIVES: Epidermal growth factor receptor (EGFR) mutation testing is a companion diagnostic to determine eligibility for treatment with EGFR tyrosine kinase inhibitors (EGFR-TKIs) in non-small cell lung cancer (NSCLC). Recently, plasma-based EGFR testing by digital polymerase chain reaction (dPCR), which enables accurate quantification of target DNA, has shown promise as a minimally invasive diagnostic. Here, we aimed to evaluate the accuracy of a plasma-based EGFR mutation test developed using chip-based dPCR-based detection of 3 EGFR mutations (exon 19 deletions, L858R in exon 21, and T790M in exon 20). MATERIALS AND METHODS: Forty-nine patients with NSCLC harboring EGFR-activating mutations were enrolled, and circulating free DNAs (cfDNAs) were extracted from the plasma of 21 and 28 patients before treatment and after progression following EGFR-TKI treatment, respectively. RESULTS: Using reference genomic DNA containing each mutation, the detection limit of each assay was determined to be 0.1%. The sensitivity and specificity of detecting exon 19 deletions and L858R mutations, calculated by comparing the mutation status in the corresponding tumors, were 70.6% and 93.3%, and 66.7% and 100%, respectively, showing similar results compared with previous studies. T790M was detected in 43% of 28 cfDNAs after progression with EGFR-TKI treatment, but in no cfDNAs before the start of the treatment. CONCLUSION: This chip-based dPCR assay can facilitate detection of EGFR mutations in cfDNA as a minimally invasive method in clinical settings.
OBJECTIVES:Epidermal growth factor receptor (EGFR) mutation testing is a companion diagnostic to determine eligibility for treatment with EGFR tyrosine kinase inhibitors (EGFR-TKIs) in non-small cell lung cancer (NSCLC). Recently, plasma-based EGFR testing by digital polymerase chain reaction (dPCR), which enables accurate quantification of target DNA, has shown promise as a minimally invasive diagnostic. Here, we aimed to evaluate the accuracy of a plasma-based EGFR mutation test developed using chip-based dPCR-based detection of 3 EGFR mutations (exon 19 deletions, L858R in exon 21, and T790M in exon 20). MATERIALS AND METHODS: Forty-nine patients with NSCLC harboring EGFR-activating mutations were enrolled, and circulating free DNAs (cfDNAs) were extracted from the plasma of 21 and 28 patients before treatment and after progression following EGFR-TKI treatment, respectively. RESULTS: Using reference genomic DNA containing each mutation, the detection limit of each assay was determined to be 0.1%. The sensitivity and specificity of detecting exon 19 deletions and L858R mutations, calculated by comparing the mutation status in the corresponding tumors, were 70.6% and 93.3%, and 66.7% and 100%, respectively, showing similar results compared with previous studies. T790M was detected in 43% of 28 cfDNAs after progression with EGFR-TKI treatment, but in no cfDNAs before the start of the treatment. CONCLUSION: This chip-based dPCR assay can facilitate detection of EGFR mutations in cfDNA as a minimally invasive method in clinical settings.
Authors: Diego Cortinovis; Umberto Malapelle; Fabio Pagni; Alessandro Russo; Giuseppe Luigi Banna; Elisa Sala; Christian Rolfo Journal: Transl Lung Cancer Res Date: 2021-07
Authors: Frederik van Delft; Hendrik Koffijberg; Valesca Retèl; Michel van den Heuvel; Maarten IJzerman Journal: Cancers (Basel) Date: 2020-04-30 Impact factor: 6.639