Gene Probe Assay of Viral Nucleic Acid Using a Silicon Biosensor,
Abstract
The use ot a silicon-based biosensor for a gene probe assay is described. The target analyte, a 391 base pair DNA fragment, was mixed with a pair of probes, one labelled with biotin, the other with fluorescein, and hybridized in homogeneous solution phase. The hybridized product was separated by biotin- streptavidin mediated filtration capture and detected using a light-addressable potentiometric sensor which monitored the presence of urease conjugated (anti-fluorescein) antibodies incorporated in the hybridized product. The total assay time, including hybridization, filtration capture and potentiometric sensing was 45 - 60 min. The lower detection limit for the assay was 0.3 fmole (1.8 x 108 molecules) of single-stranded target DNA under low stringency conditions (Tm-220C). The results indicate that the LAPS assay generates detection limits similar to conventional membrane-based colorimetric assays but in much less time. The LAPS assay is also less technically demanding.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jan 01, 1996
- Accession Number
- ADA307320
Entities
People
- Douglas E. Bader
- Glen R. Fisher
- William E. Lee
Organizations
- Defence Research and Development Canada