Development of high-speed droplet-polymerase chain reaction assay for detecting epidermal growth factor receptor gene mutation in lung cancer patient

• Project/Area Number: 25460434
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Human pathology
• Research Institution: Kyoto University (2014-2015); Shinshu University (2013)
• Principal Investigator: Yoshizawa Akihiko 京都大学, 医学(系)研究科(研究院), 講師 (80378645)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2015: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2014: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2013: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: 肺癌 / 上皮成長因子受容体変異 / 高速d-PCR法 / 新鮮細胞診材料 / 上皮成長因子受容体(EGFR) / 高速PCR法 / 高感度PCR法 / 腺癌 / EGFR / 高速化 / EGFR遺伝子変異 / KRAS遺伝子変異 / 迅速診断

Outline of Final Research Achievements

We developed a novel, rapid epidermal growth factor receptor gene (EGFR) mutation assay using a real-time droplet-polymerase chain reaction machine (EGFR d-PCR assay). The purpose of this study was to validate the performance of the EGFR d-PCR assay using fresh liquid cytology specimens (FLCSs) from lung cancer patients.　We analyzed three major EGFR mutations in 80 FLCSs via the EGFR d-PCR assay and conventional real-time PCR assay. In addition, we performed sensitivity assays using cell lines with EGFR mutations. The EGFR d-PCR assay accurately detected 3 major EGFR mutations compared to conventional real-time PCR assay, resulting in concordant between the two assays. In addition, EGFR d-PCR assay could reduce reaction time (8 min and 10 s). Sensitivity of the EGFR d-PCR assay was 0.05-0.5%. In conclusion, the EGFR d-PCR assay markedly reduced the detection time of major EGFR mutations with high sensitivity compared with the conventional assay.

Research Products

• [Presentation] 新規液滴型高速PCR装置を用いた肺腺癌EGFR遺伝子変異迅速検出法の開発 (2015)
  • Author(s): 浅香志穂、吉澤明彦 他
  • Organizer: 日本病理学会総会春季大会
  • Place of Presentation: 名古屋
  • Year and Date: 2015-04-30 – 2015-05-02
• [Presentation] 新規液滴型高速PCR装置を用いた肺腺癌EGFR遺伝子変異迅速検査法の開発 (2015)
  • Author(s): 浅香志穂, 吉澤明彦，松田和之, 山口 明美，仲田梨恵，小川薫，末木茜 ，椎名 隆之 ，張萌，小林幸弘，上原雅行，菅野光俊，本田孝行
  • Organizer: 日本病理学会
  • Place of Presentation: 名古屋市
  • Year and Date: 2015-04-30
• [Presentation] A Novel High-Speed Droplet-Polymerase Chain Reaction Can Detect Epidermal Growth Factor Receptor Gene Mutation In Non-Small Cell Lung Cancer in Less Than 10 Minutes (2015)
  • Author(s): Shiho Asaka, Akihiko Yoshizawa 他
  • Organizer: United States & Canadian Academy of Pathology
  • Place of Presentation: 米国 Boston
  • Year and Date: 2015-03-21 – 2015-03-27