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

2015 Fiscal Year Final Research Report

• 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
• 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

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.

Free Research Field

人体病理