Author of

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  P3.06 - Poster Session 3 - Prognostic and Predictive Biomarkers (ID 178)

  • Event: WCLC 2013
  • Type: Poster Session
  • Track: Biology
  • Presentations: 1
  • Moderators:
  • Coordinates: 10/30/2013, 09:30 - 16:30, Exhibit Hall, Ground Level

  • 12489

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    P3.06-049 - Detection of EGFR mutation in bronchial lavage fluid after transbronchial biopsy using a novel high-speed real-time PCR system. (ID 3246)

    09:30 - 09:30  |  Author(s): M. Takada
    • Abstract

    Background
    Epidermal growth factor receptor (EGFR) mutation testing is essential to determine treatment regimens for patients with advanced non-squamous non-small cell lung cancer (non-Sq NSCLC). However, it requires at least two weeks until the results of commercialized EGFR testing are available. Therefore, we developed a novel high-speed real-time PCR system to reduce the time required for EGFR mutation detection. The reaction time of this system is only 5 minutes, and EGFR mutation status is found out within a few hours after diagnostic bronchoscopy. We tried to detect exon 19 deletion and exon 21 point mutation(L858R) in bronchial lavage fluid (BLF) after transbronchial biopsy (TBB) using this system. The aim of this study is to assess the performance of this novel high-speed real-time PCR system.

    Methods
    Seventy five consecutive patients who underwent TBB in our hospital from November 2012 to May 2013 were enrolled. Samples were obtained from BLF after TBB. DNA was extracted using QIAamp Blood Mini Kit (QIAGEN Japan, Tokyo, Japan). EGFR mutations were detected with high-speed real-time PCR system (TRUST medical, Hyogo, Japan) (Method A). Once the patient was diagnosed NSCLC with histology of TBB samples, EGFR mutation status was validated with PCR-invader method (BML, Inc. Tokyo, Japan) (Method B). We evaluated the sensitivity and concordance between (A) and (B).

    Results
    Lung cancer was histologically diagnosed in 51 patients (adenocarcinoma/squamous cell carcinoma/others=42/6/3), while not in 24 patients. Detection rate of EGFR mutation in patients with lung cancer was 29.4%(15/51) with (A) and 31.4%(16/51) with (B), respectively. Concordance rate between two methods was 94%. Discordance was found in one (6%) sample, where minor mutation of Exon19 L747-P753 deletion and insertion S was found only with (B). When (B) was used as a standard, sensitivity and specificity of (A) were 94% and 100%, respectively. Time to detect EGFR mutation by (A) and (B) were 2 hours and 18 days (6-45 days), respectively.

    Conclusion
    A novel high-speed real-time PCR system enables us to apply rapid EGFR mutation detection for clinical use.