Detection of point mutations in cytology samples.

2019 Fiscal Year Final Research Report

• Project/Area Number: 17K15649
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Human pathology
• Research Institution: Wakayama Medical University
• Principal Investigator: Matsuzaki Ibu 和歌山県立医科大学, 医学部, 助教 (60647428)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Keywords: 細胞 / 遺伝子変異 / 細胞診

Outline of Final Research Achievements

The purpose of this study was to develop a method for analyzing gene point mutations both in vitro and in situ using cytodiagnostic materials.I examined the detection of the fusion gene, firstly.As a result, we succeeded in detecting the fusion gene of EML4-ALK, SYT-SSX, and IgH / BCL2 in vitro by the LAMP method without using the RCA method using artificial cells or cultured cells. The paper was accepted (Acta Histochem Cytochem. 2017).Since artificial nucleic acids are expensive, successful detection of gene mutations using the LAMP method alone, which is relatively inexpensive and convenient, can broaden the possibility of analyzing gene mutations using cytology samples and contribute to diagnosis and treatment.

Free Research Field

細胞診断学

Academic Significance and Societal Importance of the Research Achievements

培養細胞を用いたin vitroにて、人工核酸やRCA法は使用せず、LAMP法のみでEML4-ALK、SYT-SSX、IgH/BCL2の融合遺伝子検出に成功し、論文発表した。LAMP法は比較的安価で特別な機械を使用することなく、一定温度で反応させ、検出までの工程を１ステップで行うことができる高感度遺伝子増幅法である。また目視検出薬をあらかじめ加えて反応させることで、特別な機器を必要とせず、目視で増幅を確認できる。今後、この方法を細胞診材料に応用できれば、簡便に遺伝子解析を行うことができ、細胞診断の正診率の向上のみならず、治療戦略を立案するためのコンパニオン診断に寄与できることが期待される。