A MICROTITER PLATE ASSAY USING CASCADE AMPLIFICATION FOR DETECTION OFNONISOTOPICALLY LABELED DNA

Citation
Cb. Rothschild et al., A MICROTITER PLATE ASSAY USING CASCADE AMPLIFICATION FOR DETECTION OFNONISOTOPICALLY LABELED DNA, Analytical biochemistry, 225(1), 1995, pp. 64-72
Citations number
21
Categorie Soggetti
Biology
Journal title
ISSN journal
00032697
Volume
225
Issue
1
Year of publication
1995
Pages
64 - 72
Database
ISI
SICI code
0003-2697(1995)225:1<64:AMPAUC>2.0.ZU;2-5
Abstract
We describe a microtiter-plate-based, colorimetric assay for DNA, the enzyme-linked DNA-enzyme-linked coagulation assay (EDNA-ELCA). The EDN A-ELCA uses amplification of the common pathway of coagulation for the ultrasensitive detection of DNA which is tagged by incorporation of f unctional groups such as biotin and fluorescein. The EDNA-ELCA enables detection of attomole amounts of DNA (<1 pg per microtiter well), wit h a sensitivity 200-1000 times higher than other colorimetric techniqu es. The assay has been applied as an adjunct to PCR for quantitative d etermination of methicillin-resistant Staphylococcus aureus DNA at lev els corresponding to 1-10(5) organisms. The EDNA-ELCA can also be used to assay DNA by hybridization; <50 amol of an unlabeled DNA template is detected by hybridization to biotin- and fluorescein-labeled probes . (C) 1995 Academic Press, Inc.