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    Materials and methods for detection of nucleic acids
    1.
    发明授权
    Materials and methods for detection of nucleic acids 有权
    用于检测核酸的材料和方法

    公开(公告)号：US07517651B2

    公开(公告)日：2009-04-14

    申请号：US11486841

    申请日：2006-07-14

    申请人： David J. Marshall ,  James R. Prudent ,  Christopher B. Sherrill ,  Gideon Shapiro ,  Simon Jurczyk ,  Jerod L. Ptacin

    发明人： David J. Marshall ,  James R. Prudent ,  Christopher B. Sherrill ,  Gideon Shapiro ,  Simon Jurczyk ,  Jerod L. Ptacin

    IPC分类号： C12Q1/68

    CPC分类号： C12Q1/6853 ,  C12Q1/6823 ,  C12Q1/6827 ,  C12Q1/6832 ,  C12Q1/6834 ,  C12Q1/6858 ,  C12Q2600/156 ,  C12Q2565/1015 ,  C12Q2525/101 ,  C12Q2525/161

    摘要： Assays using non-natural bases are described. In one embodiment, the method involves contacting a sample suspected of containing the target nucleic acid with a polymerase and first and second primers; amplifying the target nucleic acid, if present in the sample, by PCR using the first and second primers to generate an amplification product having a double-stranded region and a single-stranded region that comprises the non-natural base; contacting the sample with a reporter comprising a label and a non-natural base that is complementary to the non-natural base of the single-stranded region; annealing at least a portion of the reporter to the single-stranded region of the amplification product; and correlating a signal of the label with the presence of the target nucleic acid in the sample. The invention also provides corresponding kits for use in detecting target nucleic acids in a sample.

    摘要翻译： 描述了使用非天然碱的测定法。 在一个实施方案中，所述方法包括使怀疑含有靶核酸的样品与聚合酶和第一和第二引物接触; 如果存在于样品中，则使用第一和第二引物通过PCR扩增靶核酸以产生具有双链区域和包含非天然碱基的单链区域的扩增产物; 使样品与包含与单链区域的非天然碱基互补的标记物和非天然碱基的报告子接触; 将至少一部分报告物退火至扩增产物的单链区域; 并将标记的信号与样品中靶核酸的存在相关联。 本发明还提供用于检测样品中靶核酸的相应试剂盒。

    Materials and methods for detection of nucleic acids
    3.
    发明授权
    Materials and methods for detection of nucleic acids 有权
    用于检测核酸的材料和方法

    公开(公告)号：US07422850B2

    公开(公告)日：2008-09-09

    申请号：US09861292

    申请日：2001-05-18

    申请人： David J. Marshall ,  James R. Prudent ,  Christopher B. Sherrill ,  Gideon Shapiro ,  Jennifer K. Grenier ,  Craig S. Richmond ,  Simona Jurczyk ,  Jerod L. Ptacin

    发明人： David J. Marshall ,  James R. Prudent ,  Christopher B. Sherrill ,  Gideon Shapiro ,  Jennifer K. Grenier ,  Craig S. Richmond ,  Simona Jurczyk ,  Jerod L. Ptacin

    IPC分类号： C12Q1/68

    CPC分类号： C12Q1/6853 ,  C12Q1/6823 ,  C12Q1/6827 ,  C12Q1/6832 ,  C12Q1/6834 ,  C12Q1/6858 ,  C12Q2600/156 ,  C12Q2565/1015 ,  C12Q2525/101 ,  C12Q2525/161

    摘要： Assays using non-natural bases are described. In one embodiment, the method involves contacting a sample suspected of containing the target nucleic acid with a polymerase and first and second primers; amplifying the target nucleic acid, if present in the sample, by PCR using the first and second primers to generate an amplification product having a double-stranded region and a single-stranded region that comprises the non-natural base; contacting the sample with a reporter comprising a label and a non-natural base that is complementary to the non-natural base of the single-stranded region; annealing at least a portion of the reporter to the single-stranded region of the amplification product; cleaving, after annealing, at least a portion of the reporter to release at least one reporter fragment; and correlating the release of the at least one reporter fragment with the presence of the target nucleic acid in the sample. The invention also provides corresponding kits for use in detecting target nucleic acids in a sample. Alternatively, the reporter can be incorporated into the amplification product rather than annealing and then cleaving.

    摘要翻译： 描述了使用非天然碱的测定法。 在一个实施方案中，所述方法包括使怀疑含有靶核酸的样品与聚合酶和第一和第二引物接触; 如果存在于样品中，则使用第一和第二引物通过PCR扩增靶核酸以产生具有双链区域和包含非天然碱基的单链区域的扩增产物; 使样品与包含与单链区域的非天然碱基互补的标记物和非天然碱基的报告子接触; 将至少一部分报告物退火至扩增产物的单链区域; 在退火后切割至少一部分报告物以释放至少一个报道片段; 并将至少一个报道片段的释放与样品中靶核酸的存在相关联。 本发明还提供用于检测样品中靶核酸的相应试剂盒。 或者，可以将报道子并入扩增产物，而不是退火然后裂解。