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A Microtiter Plate Assay Using Cascade Amplification for Detection of Nonisotopically Labeled DNA

✍ Scribed by C.B. Rothschild; M.X. Triscott; D.W. Bowden; G. Doellgast

Publisher
Elsevier Science
Year
1995
Tongue
English
Weight
817 KB
Volume
225
Category
Article
ISSN
0003-2697

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✦ Synopsis

We describe a microtiter-plate-based, colorimetric assay for DNA, the enzyme-linked DNA-enzymelinked coagulation assay (EDNA-ELCA). The EDNAELCA uses amplification of the common pathway of coagulation for the ultrasensitive detection of DNA which is tagged by incorporation of functional groups such as biotin and fluorescein. The EDNA-ELCA enables detection of attomole amounts of DNA ( (<1) pg per microtiter well), with a sensitivity 200-1000 times higher than other colorimetric techniques. The assay has been applied as an adjunct to PCR for quantitative determination of methicillin-resistant Staphylococcus aureus DNA at levels corresponding to (1-10^{5}) organisms. The EDNA-ELCA can also be used to assay DNA by hybridization; (<\mathbf{5 0} \mathbf{~ a m o l}) of an unlabeled DNA template is detected by hybridization to biotin- and fluorescein-labeled probes. ic 1995 Academic Press. Inc.

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