The PCR-invader method (structure-specific 5 nuclease-based method), a sensitive method for detecting EGFR gene mutations in lung cancer specimens; Comparison with direct sequencing

Katsuhiko Naoki, Kenzo Soejima, Hiroaki Okamoto, Junko Hamamoto, Naoya Hida, Ichiro Nakachi, Hiroyuki Yasuda, Sohei Nakayama, Satoshi Yoda, Ryosuke Satomi, Shinnosuke Ikemura, Hideki Terai, Takashi Sato, Koshiro Watanabe

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Abstract

Background: Several sensitive assays, including the PCR-invader method (structure-specific 5 nuclease-based method), have been used to detect EGFR mutations in non-small-cell lung cancer (NSCLC). However, validation has not been reported. We assessed the detection rate of EGFR mutation by the PCR-invader method and direct sequencing using same clinical specimens. Patients and methods: EGFR mutations were analyzed with the PCR-invader method and compared with direct sequencing using paraffin tissues and pleural and pericardial effusions from NSCLC patients. The relationships between the treatment responses and mutations were evaluated retrospectively. Results: Fifty-four samples from 42 NSCLC patients were studied. EGFR mutations were identified in 52% of the patients and 52% of the samples with the PCR-invader method, but only in 43% of the patients and in 35% of the samples by direct sequencing. In the samples obtained from the same patients at different sites and different times, EGFR mutations were coincident in nine out of ten patients by the PCR-invader method but in six out of ten patients by direct sequencing. Seventeen patients with EGFR mutations were treated with gefitinib; the response rate (RR) and disease control rate (DCR) were 41 and 94%, and median treatment duration was more than 6 months. Seven EGFR mutation-negative patients were treated with gefitinib; the RR and DCR were 0 and 14%, and median treatment duration was 1 month. Conclusion: The PCR-invader method was useful for detecting EGFR mutations in clinical lung cancer specimens and is more sensitive than direct sequencing.

Original languageEnglish
Pages (from-to)335-344
Number of pages10
JournalInternational Journal of Clinical Oncology
Volume16
Issue number4
DOIs
Publication statusPublished - 2011 Aug 1

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Keywords

  • Direct sequencing
  • EGFR mutation
  • Gefitinib
  • Non-small-cell lung cancer
  • PCR-invader method
  • Validation

ASJC Scopus subject areas

  • Surgery
  • Hematology
  • Oncology

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