A Fully Integrated Microfluidic Genetic Analysis Device for the Detection of Blood Cancers
We are developing a fully-integrated microdevice capable of DNA extraction, PCR amplification and the subsequent electrophoretic separation specifically for the detection of T-cell lymphoma (TCL). While we have recently reported a fully-integrated device for the detection of bacteria [1], the interrogation of human genomic DNA from whole blood provides new challenges, especially when detecting gene rearrangements correlative with cancer. Detection of TCL involves the PCR amplification of select sequences in the T-cell receptor gene that are likely to have undergone gene rearrangement. These PCR fragments represent a polyclonal cell population in normal individuals and a monoclonal cell population in patients with lymphoma in a way that can be discriminated by electrophoretic separation. In clinical labs, the PCR that follows DNA extraction requires ~3 hours followed by a 40 min capillary electrophoresis separation under single-stranded conditions and utilizing 4-color detection. While integration of processing steps is a large advantage over traditional methods, the reduction in the times associated with these lengthy processes is also an important benefit.
1. Easley, C.J., Karlinsey, J.M., Bienvenue, J.M., Legendre, L.A., Roper, M.G., Feldman, S.H., Hughes, M.A., Merkel, T.J., Ferrance, J.P., Landers, J.P. PNAS, 2006, 109(51), 19272-19277.
1. Easley, C.J., Karlinsey, J.M., Bienvenue, J.M., Legendre, L.A., Roper, M.G., Feldman, S.H., Hughes, M.A., Merkel, T.J., Ferrance, J.P., Landers, J.P. PNAS, 2006, 109(51), 19272-19277.
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