Collins M, Fritsch E F, Ellwood M S, Diamond S E, Williams J I, Brewen J G
Genetics Institute, Cambridge, Massachusetts.
Mol Cell Probes. 1988 Mar;2(1):15-30. doi: 10.1016/0890-8508(88)90040-0.
A novel approach for the detection of specific DNA or RNA sequences based on branch migration and DNA strand displacement is described. A partially double-stranded probe complex is prepared with a detectable label on one of the two strands and incubated with analyte molecules under hybridization conditions. The analyte molecules hybridize to the single-stranded portion of the probe complex and undergo branch migration to release the labelled DNA strand from the complex. Initial characterization of the assay indicates that both qualitative and quantitative information about analytes present in a sample can be obtained. The strand displacement assay is more sensitive to sequence alterations in the analyte than is a hybridization assay and can be promoted by rec A protein at 37 degrees C. Finally, a method for preparing probe complexes by cloning in a single-stranded DNA vector is also described.
描述了一种基于分支迁移和DNA链置换检测特定DNA或RNA序列的新方法。制备一种部分双链探针复合物，其中两条链之一带有可检测标记，并在杂交条件下与分析物分子一起孵育。分析物分子与探针复合物的单链部分杂交，并发生分支迁移以从复合物中释放标记的DNA链。该检测方法的初步表征表明，可以获得关于样品中存在的分析物的定性和定量信息。与杂交检测相比，链置换检测对分析物中的序列改变更敏感，并且可以在37℃下由Rec A蛋白促进。最后，还描述了一种通过在单链DNA载体中克隆来制备探针复合物的方法。
