Developing an Isothermal Solid-Phase Gene Mutation Testing Platform for Multiplex Gene Point Mutations Detection

Cancer is the major cause of death in Taiwan. Gene mutation is positively correlated to tumor formation. Previous researches in tumorigenesis have proved that the mutation of proto-oncogenes or tumor suppressor genes could trigger cancer formation. Currently in the art, polymerase chain reaction based methods are the main stream for mutation detection, but it not only needs expensive thermal cycler machine but also is time consuming and needs complex procedures. In this project, we try to develop an isothermal-solid phase-mutation detection platform: Solid Phase-Mutagenically Separated-Recombinase Polymerase Amplification (SP-MS-RPA). This technique apply the principle of mutagenically separated-PCR (MS-PCR), and combining both of isothermal enzymatic nucleic acid amplification reaction and isothermal hybridization reaction directly on the solid surface microarray. This SP-MS-RPA technique could reduce the steps for traditional PCR-based mutation detection methods. Nucleic acid probes that specific to target point mutation sequence were immobilized to specific position on the array and the multi-targets mutations detection were performed under isothermal condition. In this project, we also plan to collect clinical specimens from patients with/without colorectal cancer to test the clinical performance of the SP-MS-RPA technique.

Project ID:PC10507-0417
External Project ID:MOST105-2320-B182-023