A microtiter plate assay using cascade amplification for detection of nonisotopically labeled DNA.
A microtiter plate assay using cascade amplification for detection of nonisotopically labeled DNA.
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使用级联扩增检测非同位素标记 DNA 的微量滴定板测定。
DOI：
10.1006/abio.1995.1109
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发表时间：
1995
期刊：
影响因子：
--
通讯作者：
Doellgast,G
中科院分区：
文献类型：
--
作者：
Rothschild,CB;Triscott,MX;Bowden,DW;Doellgast,G
我们描述了一种基于微量平板的DNA比色法，即酶联DNA-酶联凝血试验(EDNA-ELCA)。EDNA-ELCA使用普通凝血途径的放大来超灵敏地检测DNA，该DNA通过掺入生物素和荧光素等官能团来标记。EDNA-ELCA能够检测阿托摩尔量的DNA(每微滴定1 pg)，灵敏度比其他比色技术高200-1000倍。该方法已作为聚合酶链式反应的辅助手段，用于定量检测1-105个微生物水平的耐甲氧西林金黄色葡萄球菌DNA。EDNA-ELCA还可以用于通过杂交检测DNA；通过与生物素和荧光素标记的探针杂交来检测未标记DNA模板的50amol。
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 1-105organisms. 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.