The overall goal of this project is to develop plastic, disposable, microanalysis devices capable to "on-board" PCR amplification and rapid, high resolution electrophoretic separation of DNA for rapid cancer diagnostics. Recent incidence and mortality statistics provide a substantial incentive for developing more rapid and efficient cancer diagnostics. In 1994, the number of new cancer cases approached 1,000,000 with cancer of the breast, lung and colon representing more than half of these cases. It is proposed to circumvent current cumbersome methodologies for detecting genomic alterations correlative with cancer by developing electrophoretic microanalysis "chips." Such devices have been shown to provide the same information as standard electrophoretic gels with separation times that are roughly 100-200 times faster. The main barriers preventing the employment of this technology in clinical areas such as cancer diagnostics have been the inability to effectively integrate PCR amplification into the chip platform and the difficulties associated with producing an inexpensive, disposable chip. The goals of this project aim to solve these impediments by (1) developing a novel "non-contact" approach for PCR amplification of DNA on plastic chips, and (2) defining the electrophoretic separation conditions and fluorescent tagging methodologies for rapid, high resolution separation of cancer-specific DNA fragments. These methodologies will be applied to two model cancer systems (T-cell lymphoma and colorectal cancer) to demonstrate proof of feasibility.
Thesaurus Terms: biomedical equipment development, diagnosis design /evaluation, diagnostic test, electrophoresis, neoplasm /cancer diagnosis, polymerase chain reaction DNA, colorectal neoplasm, lymphoma, temperature
