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

Research output: Contribution to journalArticle

42 Citations (Scopus)

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

Fingerprint

erbB-1 Genes
Lung Neoplasms
Polymerase Chain Reaction
Mutation
Non-Small Cell Lung Carcinoma
Pericardial Effusion
Mutation Rate
Pleural Effusion
Paraffin
Therapeutics

Keywords

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

ASJC Scopus subject areas

  • Oncology
  • Surgery
  • Hematology

Cite this

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

In: International Journal of Clinical Oncology, Vol. 16, No. 4, 08.2011, p. 335-344.

Research output: Contribution to journalArticle

Naoki, Katsuhiko ; Soejima, Kenzo ; Okamoto, Hiroaki ; Hamamoto, Junko ; Hida, Naoya ; Nakachi, Ichiro ; Yasuda, Hiroyuki ; Nakayama, Sohei ; Yoda, Satoshi ; Satomi, Ryosuke ; Ikemura, Shinnosuke ; Terai, Hideki ; Sato, Takashi ; Watanabe, Koshiro. / 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. In: International Journal of Clinical Oncology. 2011 ; Vol. 16, No. 4. pp. 335-344.
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AU - Okamoto, Hiroaki

AU - Hamamoto, Junko

AU - Hida, Naoya

AU - Nakachi, Ichiro

AU - Yasuda, Hiroyuki

AU - Nakayama, Sohei

AU - Yoda, Satoshi

AU - Satomi, Ryosuke

AU - Ikemura, Shinnosuke

AU - Terai, Hideki

AU - Sato, Takashi

AU - Watanabe, Koshiro

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N2 - 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.

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KW - EGFR mutation

KW - Gefitinib

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KW - PCR-invader method

KW - Validation

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