NUCLEIC-ACID DETECTION USING NONRADIOACTIVE LABELING METHODS
NUCLEIC-ACID DETECTION USING NONRADIOACTIVE LABELING METHODS
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DOI：
10.1006/mcpr.1995.0023
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发表时间：
1995-06-01
影响因子：
3.3
通讯作者：
FORTINA, P
中科院分区：
文献类型：
--
作者：
MANSFIELD, ES;WORLEY, JM;FORTINA, P
基于核酸探针的检测现在广泛应用于遗传研究、人类鉴定、法医学以及微生物学、血液学/肿瘤学和病毒学领域的广泛临床检测。标记探针用于各种检测格式，包括点印迹、Southern印迹（DNA靶标）、Northern印迹（RNA靶标）、Western印迹（蛋白质靶标）、原位杂交、斑块或菌落筛选以及在硅或玻璃表面固定阵列。传统上，这些检测中使用的探针具有放射性（32）磷标签，其保质期短，危险，处理成本高，并且当标记为高比活性时，可能不稳定。开发替代标记技术的广泛努力导致了比色法，化学发光和荧光分析格式。本综述总结了探针所需的特性，描述了不同的非放射性标记策略的优点和缺点，并举例说明了基于探针的检测方法，其中通过使用通用荧光扫描仪成像样品来促进检测。
Nucleic acid probe-based assays are now widely used in genetic research, human identification, forensics and in a broad spectrum of clinical assays in the fields of microbiology, haematology/oncology and virology. Labelled probes are used in a variety of assay formats including dot-blots, Southern blots (DNA target), Northern blots (RNA target), Western blots (protein target), in situ hybridization, plaque or colony screening and immobilized arrays on silicon or glass surfaces. Traditionally, the probes used in these assays have a radioactive (32)phosphorous label that has a short shelf-life, is dangerous, has high disposal costs and, when labelled to high specific-activity, may be unstable. Extensive efforts to develop alternative labelling techniques have resulted in colorimetric, chemiluminescent and fluorescent assay formats. This review summarizes the properties desired in a probe, describes the advantages and disadvantages of the different nonradioactive labelling strategies, and illustrates examples of probe-based assays in which detection is facilitated by imaging samples using a general purpose fluorescence scanner.