Prospective study of the accuracy of EGFR mutational analysis by high-resolution melting analysis in small samples obtained from patients with non-small cell lung cancer

Tomoya Fukui, Yuichiro Ohe, Koji Tsuta, Koh Furuta, Hiromi Sakamoto, Toshimi Takano, Hiroshi Nokihara, Noboru Yamamoto, Ikuo Sekine, Hideo Kunitoh, Hisao Asamura, Takaaki Tsuchida, Masahiro Kaneko, Masahiko Kusumoto, Seiichiro Yamamoto, Teruhiko Yoshida, Tomohide Tamura

Research output: Contribution to journalArticle

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Abstract

Purpose: Epidermal growth factor receptor {EGFR) mutations, especially in-frame deletions in exon 19 (DEL) and a point mutation in exon 21 (L858R), predict gefitinib sensitivity in patients with non-small cell lung cancer (NSCLC). In this study, we verified the accuracy of EGFR mutation analysis in small samples by high-resolution melting analysis (HRMA), which is a rapid method using PCR amplification with a dye to analyze the melting curves in NSCLC. Experimental Design: We designed a prospective study to compare the sensitivity and specificity of HRMA and DNA sequencing with laser capture microdissection. Eligible patients with lung lesions were screened by bronchoscopy or percutaneous needle biopsy to histologically confirm the diagnosis, followed by surgical resection of the NSCLC. Small diagnostic specimens were analyzed for EGFR mutations by HRMA, and the surgically resected specimens were examined for mutations by HRMA and DNA sequencing. Results: The analyses for EGFR mutations were conducted in 52 eligible cases of the 92 enrolled patients. EGFR mutations were detected in 18 (34.6%) patients. The results of HRMA from surgically resected specimens as well as DNA sequencing revealed 100% sensitivity and specificity. On the other hand, the sensitivity and specificity of HRMA from the small diagnostic specimens were 83.3% and 100%, respectively. Conclusions: In this study, we showed that HRMA is a highly accurate method for detecting DEL and L858R mutations in patients with NSCLC, although it is necessary to consider the identification of patients with a false-negative result when the analysis is conducted using small samples.

Original languageEnglish
Pages (from-to)4751-4757
Number of pages7
JournalClinical Cancer Research
Volume14
Issue number15
DOIs
Publication statusPublished - 2008 Aug 1
Externally publishedYes

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Epidermal Growth Factor Receptor
Non-Small Cell Lung Carcinoma
Freezing
Prospective Studies
Mutation
DNA Sequence Analysis
Sensitivity and Specificity
Exons
Laser Capture Microdissection
Needle Biopsy
Bronchoscopy
Point Mutation
Research Design
Coloring Agents
Polymerase Chain Reaction
Lung

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Prospective study of the accuracy of EGFR mutational analysis by high-resolution melting analysis in small samples obtained from patients with non-small cell lung cancer. / Fukui, Tomoya; Ohe, Yuichiro; Tsuta, Koji; Furuta, Koh; Sakamoto, Hiromi; Takano, Toshimi; Nokihara, Hiroshi; Yamamoto, Noboru; Sekine, Ikuo; Kunitoh, Hideo; Asamura, Hisao; Tsuchida, Takaaki; Kaneko, Masahiro; Kusumoto, Masahiko; Yamamoto, Seiichiro; Yoshida, Teruhiko; Tamura, Tomohide.

In: Clinical Cancer Research, Vol. 14, No. 15, 01.08.2008, p. 4751-4757.

Research output: Contribution to journalArticle

Fukui, T, Ohe, Y, Tsuta, K, Furuta, K, Sakamoto, H, Takano, T, Nokihara, H, Yamamoto, N, Sekine, I, Kunitoh, H, Asamura, H, Tsuchida, T, Kaneko, M, Kusumoto, M, Yamamoto, S, Yoshida, T & Tamura, T 2008, 'Prospective study of the accuracy of EGFR mutational analysis by high-resolution melting analysis in small samples obtained from patients with non-small cell lung cancer', Clinical Cancer Research, vol. 14, no. 15, pp. 4751-4757. https://doi.org/10.1158/1078-0432.CCR-07-5207
Fukui, Tomoya ; Ohe, Yuichiro ; Tsuta, Koji ; Furuta, Koh ; Sakamoto, Hiromi ; Takano, Toshimi ; Nokihara, Hiroshi ; Yamamoto, Noboru ; Sekine, Ikuo ; Kunitoh, Hideo ; Asamura, Hisao ; Tsuchida, Takaaki ; Kaneko, Masahiro ; Kusumoto, Masahiko ; Yamamoto, Seiichiro ; Yoshida, Teruhiko ; Tamura, Tomohide. / Prospective study of the accuracy of EGFR mutational analysis by high-resolution melting analysis in small samples obtained from patients with non-small cell lung cancer. In: Clinical Cancer Research. 2008 ; Vol. 14, No. 15. pp. 4751-4757.
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T1 - Prospective study of the accuracy of EGFR mutational analysis by high-resolution melting analysis in small samples obtained from patients with non-small cell lung cancer

AU - Fukui, Tomoya

AU - Ohe, Yuichiro

AU - Tsuta, Koji

AU - Furuta, Koh

AU - Sakamoto, Hiromi

AU - Takano, Toshimi

AU - Nokihara, Hiroshi

AU - Yamamoto, Noboru

AU - Sekine, Ikuo

AU - Kunitoh, Hideo

AU - Asamura, Hisao

AU - Tsuchida, Takaaki

AU - Kaneko, Masahiro

AU - Kusumoto, Masahiko

AU - Yamamoto, Seiichiro

AU - Yoshida, Teruhiko

AU - Tamura, Tomohide

PY - 2008/8/1

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N2 - Purpose: Epidermal growth factor receptor {EGFR) mutations, especially in-frame deletions in exon 19 (DEL) and a point mutation in exon 21 (L858R), predict gefitinib sensitivity in patients with non-small cell lung cancer (NSCLC). In this study, we verified the accuracy of EGFR mutation analysis in small samples by high-resolution melting analysis (HRMA), which is a rapid method using PCR amplification with a dye to analyze the melting curves in NSCLC. Experimental Design: We designed a prospective study to compare the sensitivity and specificity of HRMA and DNA sequencing with laser capture microdissection. Eligible patients with lung lesions were screened by bronchoscopy or percutaneous needle biopsy to histologically confirm the diagnosis, followed by surgical resection of the NSCLC. Small diagnostic specimens were analyzed for EGFR mutations by HRMA, and the surgically resected specimens were examined for mutations by HRMA and DNA sequencing. Results: The analyses for EGFR mutations were conducted in 52 eligible cases of the 92 enrolled patients. EGFR mutations were detected in 18 (34.6%) patients. The results of HRMA from surgically resected specimens as well as DNA sequencing revealed 100% sensitivity and specificity. On the other hand, the sensitivity and specificity of HRMA from the small diagnostic specimens were 83.3% and 100%, respectively. Conclusions: In this study, we showed that HRMA is a highly accurate method for detecting DEL and L858R mutations in patients with NSCLC, although it is necessary to consider the identification of patients with a false-negative result when the analysis is conducted using small samples.

AB - Purpose: Epidermal growth factor receptor {EGFR) mutations, especially in-frame deletions in exon 19 (DEL) and a point mutation in exon 21 (L858R), predict gefitinib sensitivity in patients with non-small cell lung cancer (NSCLC). In this study, we verified the accuracy of EGFR mutation analysis in small samples by high-resolution melting analysis (HRMA), which is a rapid method using PCR amplification with a dye to analyze the melting curves in NSCLC. Experimental Design: We designed a prospective study to compare the sensitivity and specificity of HRMA and DNA sequencing with laser capture microdissection. Eligible patients with lung lesions were screened by bronchoscopy or percutaneous needle biopsy to histologically confirm the diagnosis, followed by surgical resection of the NSCLC. Small diagnostic specimens were analyzed for EGFR mutations by HRMA, and the surgically resected specimens were examined for mutations by HRMA and DNA sequencing. Results: The analyses for EGFR mutations were conducted in 52 eligible cases of the 92 enrolled patients. EGFR mutations were detected in 18 (34.6%) patients. The results of HRMA from surgically resected specimens as well as DNA sequencing revealed 100% sensitivity and specificity. On the other hand, the sensitivity and specificity of HRMA from the small diagnostic specimens were 83.3% and 100%, respectively. Conclusions: In this study, we showed that HRMA is a highly accurate method for detecting DEL and L858R mutations in patients with NSCLC, although it is necessary to consider the identification of patients with a false-negative result when the analysis is conducted using small samples.

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