公开(公告)号：US20150218653A1

公开(公告)日：2015-08-06

申请号：US14424230

申请日：2013-08-16

申请人： QIAGEN GmbH

发明人： Markus Sprenger-Haussels ,  Christian Kupfer ,  Peer Schatz

IPC分类号： C12Q1/68 ,  C12Q1/70 ,  C12N15/10

CPC分类号： C12Q1/6886 ,  C12N15/101 ,  C12N15/1013 ,  C12Q1/6806 ,  C12Q1/708 ,  C12Q2563/143 ,  C12Q2600/118 ,  C12Q2600/158

摘要： A method of determining the presence or absence of a target nucleic acid in a cell sample, said method comprising: a) contacting a surface comprising anion exchange moieties with the sample under conditions suitable to induce binding between the cells and said surface; b) separating the surface with the bound cells from the remaining sample to collect the cells; c) releasing nucleic acids from the cells and d) generating a hybrid between the target nucleic acid and a probe specific for the target nucleic acid e) detecting the presence or absence of the hybrid. The present invention is provides a rapid and automatable method, wherein cells, such as epithelial cells originating from cervical swab samples, are collected from the surrounding liquid medium, such as a liquid based cytology medium, by binding them to an anion exchange surface. The cells bind directly with high affinity and quick kinetics to the anion exchange surface which preferably is provided by magnetic particles carrying anion exchange moieties. The cells that are bound to the anion exchange surface can be easily separated from the surrounding medium and can be directly resuspended in a composition that is suitable for a subsequent hybrid capturing assay which is performed in step d) to detect e. g. pathogen nucleic acids such as HPV nucleic acids.

摘要翻译： 一种确定细胞样品中靶核酸的存在或不存在的方法，所述方法包括：a）在适于诱导细胞与所述表面之间结合的条件下，使含有阴离子交换部分的表面与样品接触; b）将表面与剩余样品中的结合细胞分离以收集细胞; c）从细胞中释放核酸，以及d）在靶核酸和靶核酸特异的探针之间产生杂交体e）检测杂交体的存在或不存在。 本发明提供一种快速且可自动化的方法，其中通过将它们结合到阴离子交换表面上，从周围的液体介质如基于液体的细胞学介质中收集源自子宫颈拭子样品的细胞，例如上皮细胞。 细胞直接以高亲和力和快速动力学结合阴离子交换表面，其优选由携带阴离子交换部分的磁性颗粒提供。 与阴离子交换表面结合的细胞可以容易地与周围的培养基分离，并且可以直接重新悬浮在组合物中，该组合物适用于在步骤d）中进行的随后的杂交捕获测定，以检测e。 G。 病原体核酸如HPV核酸。