Development of High Sensitive Gene Mutation Detection Kits

Targeted therapy is an emerging therapeutic method for cancer treatment. The efficacy of targeted therapy is highly dependent on the genotypes of cancerous tissues. Therefore, companion diagnostic testing for the determination of cancer genotypes is required before treatment. The related assay kits have valuable market. The aim of this proposal is to develop companion diagnostic tests for the mutations in the genes involving in epidermal growth factor (EGFR)-related signaling pathway, including NRAS, BRAF, PIK3CA, and EGFR genes. Totally 50 types of mutations are involved. Our patented technology, peptide nucleic acid (PNA) probe, will be applied to develop ready-to-use diagnostic kits for these gene mutations. We will test different probe and primer design, optimize buffer recipe, and adjust reaction conditions, to achieve highly sensitive requirement of mutation detection, which is set at detecting 0.1-0.01% mutant alleles in the wild-type background. In addition, we will test formula to stabilize the kits at 4°C or room temperature, making them easier to handle during transportation and storage. The developed kits can be used for different types of specimens, including tissues, pleural effusions, and peripheral bloods. The detection reaction can be finished in a single tube. With these characteristics, the kits are therefore suitable for routine testing use in a clinical laboratory. In the two-year project, we plan to generate prototypes of mutation detection kits for the four genes, set their formulas, and measure their stability, sensitivity, and specificity. The kit formulas are then transfer to the corporation partner to produce the kits under Good Manufactory Protocol and seeking for further clinical trial and TFDA approval. This project not only trains qualified persons for developing and producing molecular diagnostic kits and elevates research ability of our corporation partner, but also produce kits with great market value.

Project ID:PC10505-0283
External Project ID:MOST105-2622-B182-002-CC2