公开(公告)号：US20140194311A1

公开(公告)日：2014-07-10

申请号：US14119925

申请日：2012-05-23

申请人： Mats Gullberg ,  Ola Söderberg ,  Ulf Landegren ,  Yangling Liu

发明人： Mats Gullberg ,  Ola Söderberg ,  Ulf Landegren ,  Yangling Liu

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6837 ,  C12Q1/6804 ,  C12Q1/682 ,  C12Q2531/125 ,  C12Q2533/107 ,  C12Q2537/143 ,  C12Q2563/179 ,  C12Q2565/102

摘要： The present invention provides a method for detecting interactions between or with any two of at least three target substrates, or any two of at least three features of a target substrate, or a combination of interactions and features of target substrates, by a multiplexed proximity ligation assay, said method comprising: a) for each of the at least three target substrates or features, providing a proximity probe comprising a binding moiety with affinity for the feature or binding site on said substrate, and a proximity probe oligonucleotide coupled on the binding moiety; wherein each of the proximity probe oligonucleotide carries a unique tag sequence; b) mixing the proximity probes with a sample, under a condition to allow binding of each proximity probe to its respective binding site or feature on each of said substrates through the binding moiety, c) simultaneous with, or following step b), forming circularized DNA molecules where any two proximity probes bind sufficiently close to each other on the substrate, wherein each of the circularized DNA molecules comprise complementary sequences to the unique tag sequences from the two proximity probes oligo-nucleotides; d) amplifying the circularized DNA; and e) characterizing the amplified DNA.

摘要翻译： 本发明提供了一种用于通过多路复用接近连接来检测至少三个目标衬底中的任何两个之间或与目标衬底的至少三个特征中的任何两个或目标衬底的相互作用和特征的组合之间的相互作用的方法 所述方法包括：a）对于至少三种靶基质或特征中的每一种，提供包含对所述底物上的特征或结合位点具有亲和力的结合部分的邻近探针，以及偶联在结合部分上的邻近探针寡核苷酸 ; 其中每个邻近探针寡核苷酸携带唯一的标签序列; b）将接近探针与样品混合，条件是允许通过结合部分将每个邻近探针结合到每个所述底物上的各自的结合位点或特征，c）与步骤b）同时或在步骤b）之后，形成环化的 其中任何两个接近探针在底物上彼此足够接近的DNA分子，其中每个环化的DNA分子包含来自两个邻近探针寡核苷酸的唯一标签序列的互补序列; d）扩增环化的DNA; 和e）表征扩增的DNA。