GENE PROBE ASSAY OF VIRAL NUCLEIC ACID USING A SILICON BIOSENSOR

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Authors
  1. Bader, D.E.
  2. Fisher, G.R.
  3. Lee, W.E.
Corporate Authors
Defence Research Establishment Suffield, Ralston ALTA (CAN)
Abstract
The use of 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 labeled 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 45-60min. The lower detection limit for the assay was 0.3 fmole (1.8 x 10^8) conditions (Tm-22 degrees celsius). 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.
Report Number
DRES-M-1471 — Memorandum
Date of publication
01 Oct 1995
Number of Pages
27
DSTKIM No
96-02385
CANDIS No
498219
Format(s):
Hardcopy;Hardcopy;Document Image stored on Optical Disk

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