A MICROTITER PLATE ASSAY USING CASCADE AMPLIFICATION FOR DETECTION OF NONISOTOPICALLY LABELED DNA

被引：6

作者：

ROTHSCHILD, CB

TRISCOTT, MX

BOWDEN, DW

DOELLGAST, G

机构：

[1] ELCATECH,WINSTON SALEM,NC 27101

[2] WAKE FOREST UNIV,BOWMAN GRAY SCH MED,DEPT BIOCHEM,WINSTON SALEM,NC 27157

来源：

ANALYTICAL BIOCHEMISTRY | 1995年 / 225卷 / 01期

关键词：

D O I：

10.1006/abio.1995.1109

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We describe a microtiter-plate-based, colorimetric assay for DNA, the enzyme-linked DNA-enzyme-linked coagulation assay (EDNA-ELCA). The EDNA-ELCA 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; <50 amol of an unlabeled DNA template is detected by hybridization to biotin- and fluorescein-labeled probes. (C) 1995 Academic Press, Inc.

引用

页码：64 / 72

页数：9

共 21 条

[1] AN ESTIMATE OF UNIQUE DNA-SEQUENCE HETEROZYGOSITY IN THE HUMAN GENOME [J].