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用于DNA杂交定量的玻片免疫酶测定法的改进:SIA-DNA。

Adaptation of the slide immunoenzymatic assay for quantification of DNA hybridization: SIA-DNA.

作者信息

Conway de Macario E, Jovell R J, Macario A J

机构信息

Wadsworth Center for Laboratories and Research, New York State Department of Health, Albany, NY 12201-0509.

出版信息

Biotechniques. 1990 Feb;8(2):210-7.

PMID:2180451
Abstract

A quantitative, non-radioisotopic microsystem has been developed for measuring nucleic acid hybridization using microliter volumes of test sample and reagents. This new method, Slide Immunoenzymatic Assay-DNA, is a modification of the Slide Immunoenzymatic Assay technology originally designed for quantifying antigens and antibodies. It features small, circular solid phases (circles) of transparent material for nucleic acid immobilization. This allows the use of enzyme-labelled gene probes and substrates that generate color which, due to the distribution pattern of the circles on their support, can be measured by automated microtitration plate readers. Slide Immunoenzymatic Assay-DNA has been standardized to measure hybridization of probe to purified DNA or to DNA in cells lysed directly on the circles. Owing to its simplicity, relative low cost and expeditiousness, i.e., providing results in four hours. Slide Immunoenzymatic Assay-DNA is also suitable for use in simple laboratories and field studies.

摘要

已开发出一种定量、非放射性同位素的微系统,用于使用微升体积的测试样品和试剂来测量核酸杂交。这种新方法,即玻片免疫酶法-DNA,是对最初设计用于定量抗原和抗体的玻片免疫酶法技术的改进。它具有用于核酸固定的透明材料制成的小圆形固相(圆片)。这允许使用酶标记的基因探针和产生颜色的底物,由于圆片在其载体上的分布模式,可以通过自动微量滴定板读数仪进行测量。玻片免疫酶法-DNA已标准化,用于测量探针与纯化DNA或直接在圆片上裂解的细胞中的DNA的杂交。由于其简单性、相对低成本和快速性,即在四小时内提供结果,玻片免疫酶法-DNA也适用于简单实验室和现场研究。

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