Accession Number : ADA307320
Title : Gene Probe Assay of Viral Nucleic Acid Using a Silicon Biosensor,
Corporate Author : DEFENCE RESEARCH ESTABLISHMENT SUFFIELD RALSTON (ALBERTA)
Personal Author(s) : Bader, Douglas E. ; Fisher, Glen R. ; Lee, William E.
PDF Url : ADA307320
Report Date : JAN 1996
Pagination or Media Count : 28
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.
Descriptors : *DEOXYRIBONUCLEIC ACIDS, *SILICON, *ANTIBODIES, *GENES, *HYBRIDIZATION, PROBES, DETECTION, DETECTORS, FLUORESCENCE, TIME, SOLUTIONS(GENERAL), LIMITATIONS, LOW LEVEL, HYBRID SYSTEMS, SEPARATION, BIOLOGICAL DETECTION, VIRUSES, ASSAYING, NUCLEIC ACIDS, HOMOGENEITY, FILTRATION, POTENTIOMETRIC ANALYSIS, VITAMIN B COMPLEX, UREASE.
Subject Categories : Biochemistry
Distribution Statement : APPROVED FOR PUBLIC RELEASE