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Simultaneous detection of the tumor suppressor FHIT gene and protein using the multi-functional biochip
Minoo D F Askari1, Gordon H Miller, Tuan Vo-Dinh
1Advanced Monitoring Development Group, Life Sciences Division, Graduate School of Biomedical Sciences, Oak Ridge National Laboratory & University of Tennessee/Oak Ridge, Oak Ridge, TN 37831-6101, USA. askarimd@ornl.gov
Abstract:
The tumor suppressor gene, fragile histidine triad (FHIT), encompasses the most common human chromosomal fragile site, at 3pl4.2. Detection of FHIT gene is important in cancer diagnostics since its alterations have been associated with several human cancers. A unique multi-functional biochip for simultaneous detection of FHIT DNA and FHIT protein on the same platform was applied. The design of the biochip is based on miniaturization of photodiodes, where functioning of multiple optical sensing elements, amplifiers, discriminators, and logic circuitry are integrated on a single IC board. Performance of biochip is based on biomolecular recognition processes using both DNA and protein bioreceptors, Cy5-labeled probes and laser excitation. Application of biochip for concurrent detection of various immobilized target DNA and protein molecules and multiplex of DNA and protein on the same microarray was accomplished. Linearity of biochip for quantitative measurements was demonstrated. Results demonstrated utility of this multi-functional biochip as a useful detection technology with applications in biological and clinical laboratories.
Insights
A novel biochip simultaneously detects the fragile histidine triad (FHIT) gene and protein, crucial for cancer diagnostics. This technology offers a powerful tool for biological and clinical laboratories.
Area of Science:
- Biotechnology
- Molecular Biology
- Cancer Diagnostics
Background:
- The fragile histidine triad (FHIT) gene is a tumor suppressor located at a common human chromosomal fragile site (3p14.2).
- FHIT gene alterations are linked to various human cancers, making its detection vital for cancer diagnostics.
- Current detection methods may not offer simultaneous analysis of both gene and protein levels.
Purpose of the Study:
- To develop and evaluate a unique, multi-functional biochip for the simultaneous detection of FHIT DNA and FHIT protein.
- To demonstrate the utility of this integrated platform for biological and clinical applications.
Main Methods:
- A biochip was designed integrating miniaturized photodiodes, optical sensing elements, amplifiers, discriminators, and logic circuitry on a single IC board.
- The biochip utilizes biomolecular recognition with DNA and protein bioreceptors, employing Cy5-labeled probes and laser excitation.
- Concurrent detection of immobilized target DNA and protein molecules was achieved through multiplexing on the same microarray.
Main Results:
- The multi-functional biochip successfully enabled simultaneous detection of FHIT DNA and FHIT protein.
- The biochip demonstrated linearity for quantitative measurements.
- Multiplexing of DNA and protein targets on the same microarray was accomplished.
Conclusions:
- The developed multi-functional biochip is a valuable detection technology for simultaneous FHIT DNA and protein analysis.
- This platform shows significant potential for applications in biological research and clinical diagnostics.
- The biochip offers a novel approach for advancing cancer diagnostics through comprehensive FHIT analysis.