公开(公告)号：US4818680A

公开(公告)日：1989-04-04

申请号：US811034

申请日：1985-12-18

申请人： Mary Collins ,  Joseph P. Dougherty ,  Marian S. Ellwood ,  Edward F. Fritsch ,  Kenneth A. Jacobs

发明人： Mary Collins ,  Joseph P. Dougherty ,  Marian S. Ellwood ,  Edward F. Fritsch ,  Kenneth A. Jacobs

IPC分类号： C12Q1/68 ,  G01N33/566

CPC分类号： C12Q1/6876 ,  C12Q1/6813 ,  C12Q1/6823 ,  Y10S435/81

摘要： A diagnostic reagent is disclosed which is capable of binding to a target nucleotide sequence which is bound to a labeled polynucleotide in a target binding region which is at least partially co-extensive with the target binding region in the probe polynucleotide which is capable of binding to the target nucleotide sequence. A method is disclosed in which the reagent is contacted with a sample and with a capturing polynucleotide under conditions such that target nucleotide which may be present in the sample binds to the probe polynucleotide and displaces labeled polynucleotide from the reagent complex, and the capturing polynucleotide binds selectively to the displaced labeled polynucleotide in the region of the labeled polynucleotide that had been bound to the probe polynucleotide. Determination of the displaced nucleotide gives a value which is a function of the presence and concentration of target nucleotide in the sample.

摘要翻译： 公开了一种诊断试剂，其能够结合靶向结合区域中的标记的多核苷酸的靶核苷酸序列，所述靶结合区域与探针多核苷酸中的靶结合区域至少部分共同扩增，所述靶结合区域能够结合 靶核苷酸序列。 公开了一种方法，其中试剂与样品和捕获多核苷酸在条件下接触，使得可能存在于样品中的靶核苷酸与探针多核苷酸结合并从试剂复合物置换标记的多核苷酸，并且捕获多核苷酸结合 选择性地与已经与探针多核苷酸结合的标记的多核苷酸的区域中的置换的标记的多核苷酸。 取代的核苷酸的测定给出一个值，该值是样品中靶核苷酸的存在和浓度的函数。

公开(公告)号：US4766062A

公开(公告)日：1988-08-23

申请号：US607885

申请日：1984-05-07

申请人： Steven E. Diamond ,  Joseph G. Brewen ,  Jon I. Williams ,  Marian S. Ellwood ,  Mary Collins ,  Edward F. Fritsch

发明人： Steven E. Diamond ,  Joseph G. Brewen ,  Jon I. Williams ,  Marian S. Ellwood ,  Mary Collins ,  Edward F. Fritsch

IPC分类号： G01N33/50 ,  C12N15/09 ,  C12Q1/68 ,  G01N33/58 ,  G01N33/566

CPC分类号： C12Q1/6823 ,  C12Q1/6813 ,  C12Q1/6827 ,  C12Q1/6832 ,  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）。 公开了一种与含有靶核苷酸序列（G）的样品的接触引起最初在G与靶结合区（TBR）的单链部分（IBR）之间的结合。 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）从络合物中移位。 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。

公开(公告)号：US4766064A

公开(公告)日：1988-08-23

申请号：US684308

申请日：1984-12-20

申请人： Jon I. Williams ,  Marian S. Ellwood ,  Mary Collins ,  Edward F. Fritsch ,  Joseph G. Brewen ,  Steven E. Diamond

发明人： Jon I. Williams ,  Marian S. Ellwood ,  Mary Collins ,  Edward F. Fritsch ,  Joseph G. Brewen ,  Steven E. Diamond

IPC分类号： C12Q1/68 ,  G01N33/566

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. A volume excluding polymeric agent such as poly(ethylene oxide) (PEO or PEG) or other polyethers enhances the rate of appearance of displaced labeled polynucleotide. 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）。 公开了一种与含有靶核苷酸序列（G）的样品的接触引起最初在G与靶结合区（TBR）的单链部分（IBR）之间的结合。 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）从络合物中移位。 不包括聚合物如聚（环氧乙烷）（PEO或PEG）或其他聚醚的体积增加了置换的标记多核苷酸的出现速率。 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。