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1.发明授权Detection of infectious prion protein by seeded conversion of recombinant prion protein 有权通过重组朊蛋白的种子转化检测传染性朊病毒蛋白公开(公告)号：US08216788B2
公开(公告)日：2012-07-10
申请号：US12177012
申请日：2008-07-21
IPC分类号： G01N33/53 , G01N33/48 , C12Q1/00 , C07K14/435
CPC分类号： G01N33/6896 , G01N2800/28 , G01N2800/2828
摘要： The present disclosure relates to methods and compositions for the detection of infectious proteins or prions in samples, including the diagnosis of prion related diseases. One embodiment is an ultrasensitive method for detecting PrP-res (PrPSc) that allows the use of recombinant PrP-sen (rPrP-sen) as a substrate for seeded polymerization. A sample is mixed with purified rPrP-sen to make a reaction mix which is incubated to permit aggregation of the rPrP-sen with the PrP-res that may be present in the sample. Any aggregates are intermittently disaggregated by agitation (for example by sonication) and the reaction allowed to proceed to amplify target substrate. Any rPrP-res(Sc) in the reaction mix is detected to indicate the presence of PrP-res in the original sample. This assay, which is called rPrP-PMCA, is surprisingly much faster than existing PMCA methods, yet it still retains sufficient sensitivity to detect extremely low levels of PrP-res. An alternative of rPrP-PMCA is the QUIC method in which shaking of the reaction mixture is substituted for sonication. The surprising speed and efficiency of the method permits the rapid identification and diagnosis of prion disease, which can limit the transmission of prion diseases, particularly through the food supply.
摘要翻译： 本公开涉及用于检测样品中感染性蛋白质或朊病毒的方法和组合物，包括诊断朊病毒相关疾病。 一个实施方案是用于检测允许使用重组PrP-sen（rPrP-sen）作为接种聚合的底物的PrP-res（PrPSc）的超灵敏方法。 将样品与纯化的rPrP-sen混合以制备反应混合物，将其温育以允许rPrP-sen与可能存在于样品中的PrP-res聚集。 通过搅拌（例如通过超声处理）间歇地分解任何聚集体，并使反应进行以扩增靶基质。 检测反应混合物中的任何rPrP-res（Sc）以指示原始样品中PrP-res的存在。 这种称为rPrP-PMCA的测定法比现有的PMCA方法快得多，但仍然保持足够的灵敏度来检测极低水平的PrP-res。 rPrP-PMCA的替代方案是将反应混合物的振荡代替超声处理的QUIC方法。 该方法的惊人速度和效率允许快速鉴定和诊断朊病毒疾病，这可以限制朊病毒疾病的传播，特别是通过食物供应。
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2.发明申请DETECTION OF INFECTIOUS PRION PROTEIN BY SEEDED CONVERSION OF RECOMBINANT PRION PROTEIN 审中-公开通过重组蛋白质的转化检测感染性蛋白质公开(公告)号：US20130040319A1
公开(公告)日：2013-02-14
申请号：US13489321
申请日：2012-06-05
IPC分类号： G01N33/566 , G01N21/64 , G01N33/577
CPC分类号： G01N33/6896 , G01N2800/28 , G01N2800/2828
摘要： The present disclosure relates to methods and compositions for the detection of infectious proteins or prions in samples, including the diagnosis of prion related diseases. One embodiment is an ultrasensitive method for detecting PrP-res (PrPSc) that allows the use of recombinant PrP-sen (rPrP-sen) as a substrate for seeded polymerization. A sample is mixed with purified rPrP-sen to make a reaction mix which is incubated to permit aggregation of the rPrP-sen with the PrP-res that may be present in the sample. Any aggregates are intermittently disaggregated by agitation and the reaction allowed to proceed to amplify target substrate. Any rPrP-res(Sc) in the reaction mix is detected to indicate the presence of PrP-res in the original sample. In the QUIC method in, the reaction mixture is shaken intermittently. The surprising speed and efficiency of the method permits the rapid identification and diagnosis of prion disease.
摘要翻译： 本公开涉及用于检测样品中感染性蛋白质或朊病毒的方法和组合物，包括诊断朊病毒相关疾病。 一个实施方案是用于检测允许使用重组PrP-sen（rPrP-sen）作为接种聚合的底物的PrP-res（PrPSc）的超灵敏方法。 将样品与纯化的rPrP-sen混合以制备反应混合物，将其温育以允许rPrP-sen与可能存在于样品中的PrP-res聚集。 通过搅拌间歇地分解任何聚集体，并使反应进行以扩增靶基质。 检测反应混合物中的任何rPrP-res（Sc）以指示原始样品中PrP-res的存在。 在QUIC方法中，反应混合物间歇地摇动。 该方法的惊人速度和效率允许快速鉴定和诊断朊病毒疾病。
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3.发明申请DETECTION OF INFECTIOUS PRION PROTEIN BY SEEDED CONVERSION OF RECOMBINANT PRION PROTEIN 有权通过重组蛋白质的转化检测感染性蛋白质公开(公告)号：US20090047696A1
公开(公告)日：2009-02-19
申请号：US12177012
申请日：2008-07-21
IPC分类号： C12Q1/48 , G01N33/00 , G01N33/566 , C12Q1/02
CPC分类号： G01N33/6896 , G01N2800/28 , G01N2800/2828
摘要： The present disclosure relates to methods and compositions for the detection of infectious proteins or prions in samples, including the diagnosis of prion related diseases. One embodiment is an ultrasensitive method for detecting PrP-res (PrPSc) that allows the use of recombinant PrP-sen (rPrP-sen) as a substrate for seeded polymerization. A sample is mixed with purified rPrP-sen to make a reaction mix which is incubated to permit aggregation of the rPrP-sen with the PrP-res that may be present in the sample. Any aggregates are intermittently disaggregated by agitation (for example by sonication) and the reaction allowed to proceed to amplify target substrate. Any rPrP-res(Sc) in the reaction mix is detected to indicate the presence of PrP-res in the original sample. This assay, which is called rPrP-PMCA, is surprisingly much faster than existing PMCA methods, yet it still retains sufficient sensitivity to detect extremely low levels of PrP-res. An alternative of rPrP-PMCA is the QUIC method in which shaking of the reaction mixture is substituted for sonication. The surprising speed and efficiency of the method permits the rapid identification and diagnosis of prion disease, which can limit the transmission of prion diseases, particularly through the food supply.
摘要翻译： 本公开涉及用于检测样品中感染性蛋白质或朊病毒的方法和组合物，包括诊断朊病毒相关疾病。 一个实施方案是用于检测允许使用重组PrP-sen（rPrP-sen）作为接种聚合的底物的PrP-res（PrPSc）的超灵敏方法。 将样品与纯化的rPrP-sen混合以制备反应混合物，将其温育以允许rPrP-sen与可能存在于样品中的PrP-res聚集。 通过搅拌（例如通过超声处理）间歇地分解任何聚集体，并使反应进行以扩增靶基质。 检测反应混合物中的任何rPrP-res（Sc）以指示原始样品中PrP-res的存在。 这种称为rPrP-PMCA的测定法比现有的PMCA方法快得多，但仍然保持足够的灵敏度来检测极低水平的PrP-res。 rPrP-PMCA的替代方案是将反应混合物的振荡代替超声处理的QUIC方法。 该方法的惊人速度和效率允许快速鉴定和诊断朊病毒疾病，这可以限制朊病毒疾病的传播，特别是通过食物供应。
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公开(公告)号：US06355610B2
公开(公告)日：2002-03-12
申请号：US09823494
申请日：2001-03-30
IPC分类号： A61K3800
摘要： Peptides are disclosed that inhibit the conversion of protease sensitive prion protein (PrPsen) to the protease resistant isoform (PrPres). These peptides comprise discrete fragments of prion proteins, and are shown to inhibit the in vitro conversion of PrPsen to PrPres in a cell-free system. Numerous peptides are disclosed that include at least two amino acid residues from the highly amyloidogenic region P113-120 of the PrP protein. None of these peptides conferred protease-resistance to the PrPsen molecules. The presence of residues 119 and 120 from the highly hydrophobic sequence AGAAAAGA (position 113 to 120) produced a particular inhibitory effect. The inhibition occurred with a high degree of specificity (e.g. with an IC50 of less than about 1000 &mgr;M).
摘要翻译： 公开了抑制蛋白酶敏感性朊病毒蛋白（PrPsen）转化为蛋白酶抗性同种型（PrPres）的肽。 这些肽包含朊病毒蛋白的离散碎片，并且显示出在无细胞系统中抑制PrPsen到PrPres的体外转化。 公开了许多肽，其包括来自PrP蛋白质的高淀粉样蛋白原性区域P113-120的至少两个氨基酸残基。 这些肽都没有赋予PrPsen分子蛋白酶抗性。 来自高疏水性序列AGAAAAGA（113至120位）的残基119和120的存在产生特别的抑制作用。 抑制发生在高度的特异性（例如，IC 50小于约1000μM）。
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公开(公告)号：US20130288389A1
公开(公告)日：2013-10-31
申请号：US13979843
申请日：2012-01-17
IPC分类号： G01N33/68
CPC分类号： G01N33/6896 , G01N2800/2828
摘要： Methods are disclosed for detecting prions and/or prion disease-associated forms of prion protein. These methods provide sensitive and specific identification of prions in both biological and environmental samples. These methods include the use of both immunoprecipitation and an amplification assay that uses shaking in the absence of sonication, such as QuIC(SQ) or RT-QuIC(RTQ). In specific non-limiting examples, the methods include the use of monoclonal antibody 15B3 and/or RT-QuIC(RTQ), and/or a substrate replacement step.
摘要翻译： 公开了用于检测朊病毒蛋白和/或朊病毒疾病相关形式的朊病毒蛋白的方法。 这些方法提供生物和环境样品中朊病毒的敏感和特异性鉴定。 这些方法包括使用免疫沉淀和在不存在超声处理的情况下使用振荡的扩增测定法，例如QuIC（SQ）或RT-QuIC（RTQ）。 在具体的非限制性实例中，所述方法包括使用单克隆抗体15B3和/或RT-QuIC（RTQ），和/或使用底物置换步骤。
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公开(公告)号：US06211149B1
公开(公告)日：2001-04-03
申请号：US09128450
申请日：1998-08-03
IPC分类号： A61K3800
摘要： Peptides are disclosed that inhibit the conversion of protease sensitive prion protein (PrPsen) to the protease resistant isoform (PrPres). These peptides comprise discrete fragments of prion proteins, and are shown to inhibit the in vitro conversion of PrPsen to PrPres in a cell-free system. Numerous peptides are disclosed that include at least two amino acid residues from the highly amyloidogenic region P113-120 of the PrP protein. None of these peptides conferred protease-resistance to the PrPsen molecules. The presence of residues 119 and 120 from the highly hydrophobic sequence AGAAAAGA (position 113 to 120) produced a particular inhibitory effect. The inhibition occurred with a high degree of specificity (e.g. with an IC50 of less than about 1000 &mgr;M).
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