A novel, rapid point-of-care test for lung cancer patients to detect epidermal growth factor receptor gene mutations by using real-time droplet-PCR and fresh liquid cytology specimens

  • Authors:
    • Shiho Asaka
    • Akihiko Yoshizawa
    • Kazuyuki Matsuda
    • Akemi Yamaguchi
    • Hiroshi Yamamoto
    • Takayuki Shiina
    • Rie Nakata
    • Kaoru Ogawa
    • Meng Zhang
    • Takayuki Honda
  • View Affiliations

  • Published online on: December 2, 2016     https://doi.org/10.3892/or.2016.5287
  • Pages: 1020-1026
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Abstract

Epidermal growth factor receptor gene (EGFR) mutations are associated with response to tyrosine kinase inhibitors (TKIs) in patients with non-small cell lung cancer (NSCLC). We developed a novel, rapid EGFR mutation assay using a real-time droplet-polymerase chain reaction machine (EGFR d-PCR assay). The purpose of this study was to validate the performance of the EGFR d-PCR assay using fresh liquid cytology specimens. We analyzed three major EGFR mutations (L858R in exon 21, E746_A750del in exon 19 and T790M in exon 20) in 80 fresh liquid cytology specimens of adenocarcinoma (ADC) or NSCLC-not otherwise specified (NOS) via the EGFR d-PCR assay and conventional real-time PCR assay using the therascreen® EGFR RGQ PCR kit (Therascreen assay). In addition, we performed sensitivity assays using cell lines with EGFR mutations. The EGFR d-PCR assay detected 16 L858Rs, 8 E746_A750dels and 1 T790M mutation and the Therascreen assay detected 16 L858Rs, 11 deletions in exon 19 and 1 T790M mutation. The results were concordant between the two assays. The reaction time of the EGFR d-PCR assay was 8 min and 10 sec, but that of the Therascreen assay was 1 h and 45 min. Sensitivity, as assessed by the detection limit of the EGFR d-PCR assay was 0.5, 0.05 and 0.5% for L858R, E746_A750del and T790M, respectively. The EGFR d-PCR assay markedly reduced the detection time of major EGFR mutations with high sensitivity compared with the conventional Therascreen assay and is expected to expedite EGFR-TKI therapy for lung cancer patients, especially those in advanced stages.
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February-2017
Volume 37 Issue 2

Print ISSN: 1021-335X
Online ISSN:1791-2431

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Spandidos Publications style
Asaka S, Yoshizawa A, Matsuda K, Yamaguchi A, Yamamoto H, Shiina T, Nakata R, Ogawa K, Zhang M, Honda T, Honda T, et al: A novel, rapid point-of-care test for lung cancer patients to detect epidermal growth factor receptor gene mutations by using real-time droplet-PCR and fresh liquid cytology specimens. Oncol Rep 37: 1020-1026, 2017.
APA
Asaka, S., Yoshizawa, A., Matsuda, K., Yamaguchi, A., Yamamoto, H., Shiina, T. ... Honda, T. (2017). A novel, rapid point-of-care test for lung cancer patients to detect epidermal growth factor receptor gene mutations by using real-time droplet-PCR and fresh liquid cytology specimens. Oncology Reports, 37, 1020-1026. https://doi.org/10.3892/or.2016.5287
MLA
Asaka, S., Yoshizawa, A., Matsuda, K., Yamaguchi, A., Yamamoto, H., Shiina, T., Nakata, R., Ogawa, K., Zhang, M., Honda, T."A novel, rapid point-of-care test for lung cancer patients to detect epidermal growth factor receptor gene mutations by using real-time droplet-PCR and fresh liquid cytology specimens". Oncology Reports 37.2 (2017): 1020-1026.
Chicago
Asaka, S., Yoshizawa, A., Matsuda, K., Yamaguchi, A., Yamamoto, H., Shiina, T., Nakata, R., Ogawa, K., Zhang, M., Honda, T."A novel, rapid point-of-care test for lung cancer patients to detect epidermal growth factor receptor gene mutations by using real-time droplet-PCR and fresh liquid cytology specimens". Oncology Reports 37, no. 2 (2017): 1020-1026. https://doi.org/10.3892/or.2016.5287