Usefulness of peptide nucleic acid (PNA)-clamp smart amplification process version 2 (SmartAmp2) for clinical diagnosis of KRAS codon12 mutations in lung adenocarcinoma: Comparison of PNA-clamp SmartAmp2 and PCR-related methods

Takuya Araki, Kimihiro Shimizu, Katsunori Nakamura, Tomonori Nakamura, Yasumasa Mitani, Kyoko Obayashi, Yukiyoshi Fujita, Seiichi Kakegawa, Yohei Miyamae, Kyoichi Kaira, Takefumi Ishidao, Alexander Lezhava, Yoshihide Hayashizaki, Izumi Takeyoshi, Koujirou Yamamoto

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

KRAS is an oncogene that can be activated by mutations. Patients with non-small cell lung cancer who have KRAS mutations do not respond to tyrosine kinase inhibitors; therefore, accurate detection of KRAS mutations is important for deciding therapeutic strategies. Although sequencing-related techniques have been frequently used, they are usually too complex, have low sensitivity, and are time-consuming for routine screening in clinical situations. We evaluated peptide nucleic acid (PNA)-clamp smart amplification process version 2 (SmartAmp2) as a detection method for KRAS codon 12 mutations in patient specimens compared with traditional sequencing and polymerase chain reaction-related methods. Among 172 lung adenocarcinoma samples, direct sequencing, enzyme-enriched sequencing, and PNA-enriched sequencing showed that 16 (9.3%), 26 (15.7%), and 28 (16.3%) tumors, respectively, contained KRAS mutations in codon 12. Using PNA-clamp SmartAmp2, we could identify 31 (18.0%) tumors that had KRAS mutations in codon 12 within 60 minutes, three of which were undetected by polymerase chain reaction-related methods. On the other hand, we examined 30 nonmalignant peripheral lung tissue specimens and found no mutations in any of the samples using PNA-clamp Smart-Amp2. In this study, we confirmed that PNA-clamp Smart-Amp2 has high sensitivity and accuracy and is suitable for the clinical diagnosis of KRAS codon 12 mutations.

Original languageEnglish
Pages (from-to)118-124
Number of pages7
JournalJournal of Molecular Diagnostics
Volume12
Issue number1
DOIs
Publication statusPublished - 2010 Jan
Externally publishedYes

Fingerprint

Peptide Nucleic Acids
Polymerase Chain Reaction
Mutation
Codon
Adenocarcinoma of lung
Oncogenes
Non-Small Cell Lung Carcinoma
Protein-Tyrosine Kinases
Neoplasms
Lung

ASJC Scopus subject areas

  • Molecular Medicine
  • Pathology and Forensic Medicine

Cite this

Usefulness of peptide nucleic acid (PNA)-clamp smart amplification process version 2 (SmartAmp2) for clinical diagnosis of KRAS codon12 mutations in lung adenocarcinoma : Comparison of PNA-clamp SmartAmp2 and PCR-related methods. / Araki, Takuya; Shimizu, Kimihiro; Nakamura, Katsunori; Nakamura, Tomonori; Mitani, Yasumasa; Obayashi, Kyoko; Fujita, Yukiyoshi; Kakegawa, Seiichi; Miyamae, Yohei; Kaira, Kyoichi; Ishidao, Takefumi; Lezhava, Alexander; Hayashizaki, Yoshihide; Takeyoshi, Izumi; Yamamoto, Koujirou.

In: Journal of Molecular Diagnostics, Vol. 12, No. 1, 01.2010, p. 118-124.

Research output: Contribution to journalArticle

Araki, T, Shimizu, K, Nakamura, K, Nakamura, T, Mitani, Y, Obayashi, K, Fujita, Y, Kakegawa, S, Miyamae, Y, Kaira, K, Ishidao, T, Lezhava, A, Hayashizaki, Y, Takeyoshi, I & Yamamoto, K 2010, 'Usefulness of peptide nucleic acid (PNA)-clamp smart amplification process version 2 (SmartAmp2) for clinical diagnosis of KRAS codon12 mutations in lung adenocarcinoma: Comparison of PNA-clamp SmartAmp2 and PCR-related methods', Journal of Molecular Diagnostics, vol. 12, no. 1, pp. 118-124. https://doi.org/10.2353/jmoldx.2010.090081
Araki, Takuya ; Shimizu, Kimihiro ; Nakamura, Katsunori ; Nakamura, Tomonori ; Mitani, Yasumasa ; Obayashi, Kyoko ; Fujita, Yukiyoshi ; Kakegawa, Seiichi ; Miyamae, Yohei ; Kaira, Kyoichi ; Ishidao, Takefumi ; Lezhava, Alexander ; Hayashizaki, Yoshihide ; Takeyoshi, Izumi ; Yamamoto, Koujirou. / Usefulness of peptide nucleic acid (PNA)-clamp smart amplification process version 2 (SmartAmp2) for clinical diagnosis of KRAS codon12 mutations in lung adenocarcinoma : Comparison of PNA-clamp SmartAmp2 and PCR-related methods. In: Journal of Molecular Diagnostics. 2010 ; Vol. 12, No. 1. pp. 118-124.
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abstract = "KRAS is an oncogene that can be activated by mutations. Patients with non-small cell lung cancer who have KRAS mutations do not respond to tyrosine kinase inhibitors; therefore, accurate detection of KRAS mutations is important for deciding therapeutic strategies. Although sequencing-related techniques have been frequently used, they are usually too complex, have low sensitivity, and are time-consuming for routine screening in clinical situations. We evaluated peptide nucleic acid (PNA)-clamp smart amplification process version 2 (SmartAmp2) as a detection method for KRAS codon 12 mutations in patient specimens compared with traditional sequencing and polymerase chain reaction-related methods. Among 172 lung adenocarcinoma samples, direct sequencing, enzyme-enriched sequencing, and PNA-enriched sequencing showed that 16 (9.3{\%}), 26 (15.7{\%}), and 28 (16.3{\%}) tumors, respectively, contained KRAS mutations in codon 12. Using PNA-clamp SmartAmp2, we could identify 31 (18.0{\%}) tumors that had KRAS mutations in codon 12 within 60 minutes, three of which were undetected by polymerase chain reaction-related methods. On the other hand, we examined 30 nonmalignant peripheral lung tissue specimens and found no mutations in any of the samples using PNA-clamp Smart-Amp2. In this study, we confirmed that PNA-clamp Smart-Amp2 has high sensitivity and accuracy and is suitable for the clinical diagnosis of KRAS codon 12 mutations.",
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AU - Obayashi, Kyoko

AU - Fujita, Yukiyoshi

AU - Kakegawa, Seiichi

AU - Miyamae, Yohei

AU - Kaira, Kyoichi

AU - Ishidao, Takefumi

AU - Lezhava, Alexander

AU - Hayashizaki, Yoshihide

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