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Simultaneous Detection
of the FHIT Gene and Protein Using Microarray-based Biochip
Askari, Minoo1,2,
Miller, Gordon1 and Vo-Dinh, Tuan1,2
Oak Ridge National Laboratory1
University of Tennessee/Knoxville2
Abstract-
The tumor suppressor gene, FHIT, fragile histadine triad, encompasses
the most common human chromosomal fragile site, at 3p14.2. Detection
of the FHIT tumor suppressor gene is important in cancer diagnostics
since alterations in this gene have been associated with carcinogenic
manifestations in several human cancers. In this work we used a unique
multifuntional biochip for simultaneous detection of FHIT DNA and FHIT
protein extracted from invitro cell cultures and actual mice tumors,
on the same platform. The biochip system is based on miniaturization
of phototransistors, where the functioning of multiple optical sensing
elements, amplifiers, discriminators, and logic circuitry are integrated
on a single IC board. The performance of the biochip is based on biomolecular
recognition processes using both DNA and protein as bioreceptors. The
detection system utilizes Cy5 dye labeled probes and laser excitation
to detect both FHIT protein and FHIT DNA immobilized on a nylon membrane
platform. The performance of the integrated phototransistors and amplifier
circuits of the biochip, analyzed through measurement of the signal
output response for various concentrations of fluorescently-labeled
DNA and protein molecules, have illustrated the linearity of the biochip
essential for quantitative and qualitative target detection at very
low concentrations. These features demonstrate the utility of this biochip
as a simple yet sensitive and selective tool for cancer detection in
a biological research laboratory and in clinical settings.
Keywords: FHIT,
Fragile Histidine Triad gene, Biochip, Fluorescence detection technology,
Microarray technology
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