公开(公告)号：US07932060B2

公开(公告)日：2011-04-26

申请号：US10826654

申请日：2004-04-19

申请人： James Nadeau ,  Tobin Hellyer ,  Dolores Berger ,  William Nussbaumer ,  Robert Rosenstein ,  Andrew Kuhn ,  Sha Sha Wang ,  Keith Thornton

发明人： James Nadeau ,  Tobin Hellyer ,  Dolores Berger ,  William Nussbaumer ,  Robert Rosenstein ,  Andrew Kuhn ,  Sha Sha Wang ,  Keith Thornton

IPC分类号： C12P19/34 ,  C12Q1/68 ,  G01N33/53 ,  C07H21/04 ,  C07K1/00

CPC分类号： C12Q1/6804 ,  C12Q1/6851

摘要： A high-sensitivity, low-background immuno-amplification assay is provided, which offers a streamlined workflow suitable for high-throughput assays of clinically relevant samples, such as blood and other bodily fluids. The assay comprises the use of two proximity members that each comprise an analyte-specific binding component conjugated to an oligonucleotide. Binding an analyte brings the oligonucleotide moieties of the proximity members in sufficiently close contact that the oligonucleotides form an amplicon. The presence of the analyte then is detected through amplification of the amplicon and detection of the amplified nucleic acids. The sensitivity of the assay of the present invention is improved by preventing spurious or non-specific amplicon formation by proximity members that are not complexed with an analyte.

摘要翻译： 提供了高灵敏度，低背景的免疫扩增测定法，其提供流线型工作流程，适用于临床相关样品如血液和其他体液的高通量测定。 该测定包括使用两个邻近成员，每个邻近成员包含与寡核苷酸缀合的分析物特异性结合成分。 结合分析物使邻近成员的寡核苷酸部分充分紧密接触，使寡核苷酸形成扩增子。 然后通过扩增扩增子和扩增核酸的检测来检测分析物的存在。 通过防止与分析物不复合的邻近成员形成假的或非特异性的扩增子来提高本发明的测定的灵敏度。

公开(公告)号：US20050009050A1

公开(公告)日：2005-01-13

申请号：US10826654

申请日：2004-04-19

申请人： James Nadeau ,  Tobin Hellyer ,  Dolores Berger ,  William Nussbaumer ,  Robert Rosenstein ,  Andrew Kuhn ,  Sha Wang ,  Keith Thornton

发明人： James Nadeau ,  Tobin Hellyer ,  Dolores Berger ,  William Nussbaumer ,  Robert Rosenstein ,  Andrew Kuhn ,  Sha Wang ,  Keith Thornton

IPC分类号： C07K20060101 ,  C12P19/34 ,  C12Q1/68

CPC分类号： C12Q1/6804 ,  C12Q1/6851

摘要： A high-sensitivity, low-background immuno-amplification assay is provided, which offers a streamlined workflow suitable for high-throughput assays of clinically relevant samples, such as blood and other bodily fluids. The assay comprises the use of two proximity members that each comprise an analyte-specific binding component conjugated to an oligonucleotide. Binding an analyte brings the oligonucleotide moieties of the proximity members in sufficiently close contact that the oligonucleotides form an amplicon. The presence of the analyte then is detected through amplification of the amplicon and detection of the amplified nucleic acids. The sensitivity of the assay of the present invention is improved by preventing spurious or non-specific amplicon formation by proximity members that are not complexed with an analyte. In one embodiment, target-independent amplicon formation is prevented by using hybridization blocker oligonucleotides that bind oligonucleotide moieties that are not hybridized to each other. Background is further reduced by providing a solid phase capture oligonucleotide that prevents amplicon formation until the captured complex is released.

摘要翻译： 提供了高灵敏度，低背景的免疫扩增测定法，其提供流线型工作流程，适用于临床相关样品如血液和其他体液的高通量测定。 该测定包括使用两个邻近成员，每个邻近成员包含与寡核苷酸缀合的分析物特异性结合成分。 结合分析物使邻近成员的寡核苷酸部分充分紧密接触，使寡核苷酸形成扩增子。 然后通过扩增扩增子和扩增核酸的检测来检测分析物的存在。 通过防止与分析物不复合的邻近成员形成假的或非特异性的扩增子来提高本发明的测定的灵敏度。 在一个实施方案中，通过使用结合彼此不杂交的寡核苷酸部分的杂交阻断剂寡核苷酸来防止靶标无关扩增子形成。 通过提供一种固相捕获寡核苷酸来进一步减少背景，所述寡核苷酸防止扩增子形成，直到所捕获的复合物被释放。