公开(公告)号：EP0164876B1

公开(公告)日：1988-03-23

申请号：EP85303176.3

申请日：1985-05-03

申请人： ALLIED CORPORATION ,  GENETICS INSTITUTE, INC.

发明人： Collins, Mary ,  Fritsch, Edward Francis ,  Williams, Jon Ira ,  Brewen, Joseph Grant ,  Diamond, Steven Elliot ,  Ellwood, Marian Sue

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6823 ,  C12Q1/6827 ,  C12Q1/6832 ,  C12Q1/6876

公开(公告)号：EP0219842A1

公开(公告)日：1987-04-29

申请号：EP86114509.2

申请日：1986-10-20

申请人： ALLIED CORPORATION ,  GENETICS INSTITUTE, INC.

发明人： Ellwood, Marian Sue ,  Collins, Mary ,  Diamond, Steven Elliot

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6832 ,  C12Q1/6813

摘要： A method for determining the presence of a predetermined target nucleotide sequence in the nucleic acid of a biological sample which comprises the steps:

(a) contacting, in solution, the sample and (i) a labeled first probe polynucleotide being capable of base pair binding to the target nucleotide sequence and (ii) an immobilizable or immobilized second probe polynucleotide which is capable of base pair binding selectively to the first probe polynucleotide in a region of first probe polynucleotide capable of base pair binding to the target nucleotide sequence,
under conditions in which first probe polynucleotide can bind to target nucleotide sequence and can bind to second probe polynucleotide,
(b) immobilizing the second probe polynucleotide, if immobilizable, with any labeled first probe polynucleotide bound thereto, and
(c) detecting the labeled polynucleotide which is not immobilized with the immobilized or immobilizable second probe polynucleotide as a measure of the amount of target nucleotide sequence in the sample.

Also, a kit comprising such a labeled first polynucleotide and immobilizable second polynucleotide and immobilization means for immobilizing the immobilizable second polynucleotide together with any labeled first polynucleotide bound thereto.
Also, a similar method or kit wherein the first probe polynucleotide is immobilizable or immobilized and the second probe polynucleotide is labeled.

摘要翻译： 一种确定生物样品核酸中预定靶核苷酸序列的存在的方法，其包括以下步骤：（a）在溶液中与样品接触，和（i）能够与碱基对结合的标记的第一探针多核苷酸 到目标核苷酸序列和（ii）可固定或固定的第二探针多核苷酸，其能够在能够碱基对结合靶核苷酸序列的第一探针多核苷酸的区域中选择性地选择性地结合第一探针多核苷酸， 所述第一探针多核苷酸可以结合靶核苷酸序列并且可以结合第二探针多核苷酸，（b）将固定第二探针多核苷酸（如果可固定）与任何与其结合的任何标记的第一探针多核苷酸固定，以及（c） 用固定的或可固定的第二探针多核苷酸固定，作为焦油量的量度 得到样品中的核苷酸序列。 另外，包含这种标记的第一多核苷酸和可固定的第二多核苷酸的试剂盒和用于将可固定的第二多核苷酸与任何与其结合的任何标记的第一多核苷酸固定的固定化装置。 此外，类似的方法或试剂盒，其中第一探针多核苷酸是可固定的或固定的，第二探针多核苷酸被标记。

公开(公告)号：EP0167238B1

公开(公告)日：1988-06-01

申请号：EP85303175.5

申请日：1985-05-03

申请人： ALLIED CORPORATION ,  GENETICS INSTITUTE, INC.

发明人： Collins, Mary ,  Fritsch, Edward Francis ,  Williams, Jon Ira ,  Brewen, Joseph Grant ,  Diamond, Steven Elliot ,  Ellwood, Marian Sue

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6832 ,  C12Q1/6823 ,  C12Q1/6827 ,  C12Q1/6876 ,  Y10S435/803 ,  Y10S435/81

公开(公告)号：EP0164876A1

公开(公告)日：1985-12-18

申请号：EP85303176.3

申请日：1985-05-03

申请人： ALLIED CORPORATION ,  GENETICS INSTITUTE, INC.

发明人： Collins, Mary ,  Fritsch, Edward Francis ,  Williams, Jon Ira ,  Brewen, Joseph Grant ,  Diamond, Steven Elliot ,  Ellwood, Marian Sue

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6823 ,  C12Q1/6827 ,  C12Q1/6832 ,  C12Q1/6876

摘要： A diagnostic reagent is disclosed containing a complex of a probe polynucleotide (P) bound via purine/pyrimidine hydrogen bonding to a labeled polynucleotide (L). The probe (P) contains a target binding region (TBR) capable of binding to a target nucleotide sequence (G) of a biological sample. A method is disclosed in which contact with a sample containing the target nucleotide sequence (G) in the presence of a recombination protein and its cofactors causes binding, initially between G and a single-stranded portion (IBR) of the target binding region (TBR). Thereafter the labeled polynucleotide (L) is displaced from the complex by branch migration of (G) into the (P)/(L) binding region. Determination of displaced labeled polynucleotide (L) gives a value which is a function of the presence and concentration of target nucleotide sequence (G) in the sample. The presence of the recombination protein (such as the rec A protein from the enteric bacterium E. coli) enhances the rate of appearance of displaced labeled polynucleotide, especially when the reaction is conducted at or near physiological temperatures

摘要翻译： 公开了一种诊断试剂，其含有通过嘌呤/嘧啶氢键与标记的多核苷酸（L）结合的探针多核苷酸（P）的复合物。 探针（P）含有能够结合生物样品的靶核苷酸序列（G）的靶结合区（TBR）。 公开了一种方法，其中在存在重组蛋白及其辅因子的情况下与含有靶核苷酸序列（G）的样品的接触引起最初在G与目标结合区的单链部分（IBR）之间的结合（TBR ）。 此后，通过（G）的分支迁移进入（P）/（L）结合区域，标记的多核苷酸（L）从配合物中移位。 置换的标记多核苷酸（L）的测定给出了样品中靶核苷酸序列（G）的存在和浓度的函数值。 重组蛋白（例如来自肠杆菌大肠杆菌的rec A蛋白）的存在增加了置换的标记多核苷酸的出现速率，特别是当反应在生理温度或接近生理温度下进行时

公开(公告)号：EP0167238A1

公开(公告)日：1986-01-08

申请号：EP85303175.5

申请日：1985-05-03

申请人： ALLIED CORPORATION ,  GENETICS INSTITUTE, INC.

发明人： Collins, Mary ,  Fritsch, Edward Francis ,  Williams, Jon Ira ,  Brewen, Joseph Grant ,  Diamond, Steven Elliot ,  Ellwood, Marian Sue

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6832 ,  C12Q1/6823 ,  C12Q1/6827 ,  C12Q1/6876 ,  Y10S435/803 ,  Y10S435/81

摘要： A diagnostic reagent is disclosed containing a complex of a probe polynucleotide (P) bound via purine/pyrimidine hydrogen bonding to a labeled polynucleotide (L). The probe (P) contains a target binding region (TBR) capable of binding to a target nucleotide sequence (G) of a biological sample. A method is disclosed in which contact with a sample containing the target nucleotide sequence (G) causes binding, initially between G and a single-stranded portion (IBR) of the target binding region (TBR). Thereafter the labeled polynucleotide (L) is displaced from the complex by branch migration of (G) into the (P)/(L) binding region. Determination of displaced labeled polynucleotide (L) gives a value which is a function of the presence and concentration of target nucleotide sequence (G) in the sample.

摘要翻译： 公开了一种诊断试剂，其含有通过与标记的多核苷酸（L）氢键结合的嘌呤/嘧啶结合的探针多核苷酸（P）的复合物。 探针（P）包含能够结合生物样品的靶核苷酸序列（G）的靶结合区域（TBR）。 公开了一种方法，其中与包含靶核苷酸序列（G）的样品的接触初始在G与靶结合区域（TBR）的单链部分（IBR）之间引起结合。 此后，标记的多核苷酸（L）通过（G）的分支迁移到（P）/（L）结合区域从复合体中置换。 置换标记多核苷酸（L）的测定给出的值是样品中靶核苷酸序列（G）的存在和浓度的函数。