核酸杂交:从研究工具到常规诊断方法。

Medical biology Pub Date : 1986-01-01
A C Syvänen
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引用次数: 0

摘要

核酸杂交反应具有极强的特异性,因此是鉴定感兴趣的基因或生物体的一种有价值的工具。随着核酸杂交技术在诊断医学等应用领域的应用越来越广泛,已经开发出比原来用于基础研究的更方便的杂交检测方法。在传统的核酸杂交方法中,固定化的核酸是通过放射性标记探针在过滤器上检测的。夹层杂交是一种简单的测试形式,用于分析未纯化的生物材料,但缺点是反应速度慢。如果有一种方法可以从反应混合物中分离形成的杂化物,则溶液杂化方法快速且容易执行。在非同位素检测中,核酸探针用化学基团修饰,经过杂交后与标记的检测器分子鉴定。检测灵敏度低是目前核酸杂交方法存在的主要问题。放大可检测信号或可检测核酸序列数量的程序是解决这一问题的潜在方法。新的杂交方法已经成功地用于一些应用,但仍然需要结合到执行良好的测试,以适用于任何期望的目的。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Nucleic acid hybridization: from research tool to routine diagnostic method.

The nucleic acid hybridization reaction is extremely specific and thus a valuable tool for the identification of genes or organism of interest. The increasing use of nucleic acid hybridization in applied fields like diagnostic medicine has led to the development of more convenient hybridization assays than those originally used in basic research. In conventional nucleic acid hybridization methods immobilized nucleic acids are detected on a filter by a radiolabelled probe. Sandwich hybridization is a simple test format for the analysis of unpurified biological material, but has the disadvantage of a slow reaction rate. Solution hybridization methods are fast and easy to perform provided that a method to separate the formed hybrids from the reaction mixture is available. In non-isotopic detection the nucleic acid probe is modified with a chemical group, which is identified with a labelled detector molecule after hybridization. The low sensitivity of detection is the main problem in nucleic acid hybridization methods. Procedures to amplify the detectable signal or the amount of detectable nucleic acid sequences are potential solutions to this problem. The new hybridization methods have successfully been used for some applications, but still need to be combined into well performing tests to be applicable to any desired purpose.

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