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公开(公告)号：US5710264A
公开(公告)日：1998-01-20
申请号：US478085
申请日：1995-06-07
申请人： Michael S. Urdea , Thomas Horn , Chu-An Chang , Brian Warner , Timothy J. Fultz
发明人： Michael S. Urdea , Thomas Horn , Chu-An Chang , Brian Warner , Timothy J. Fultz
摘要： Large comb-type branched polynucleotides useful as amplifier molecules in nucleic acid hybridization assays are provided, the polynucleotides comprising: a polynucleotide backbone having at least about 15 multifunctional nucleotides, each of which defines a sidechain site and a first single-stranded oligonucleotide unit that is capable of binding specifically to a first single-stranded polynucleotide sequence of interest; and pendant polynucleotide sidechains extending from said multifunctional nucleotides each comprising iterations of a second single-stranded oligonucleotide unit that is capable of binding specifically to a second single-stranded polynucleotide sequence of interest, the total number of iterations in all sidechains being at least 20. Methods of making these polynucleotides are also provided.
摘要翻译： 提供了用作核酸杂交测定中的放大分子的大梳状支化多核苷酸，所述多核苷酸包含：多核苷酸主链，其具有至少约15个多功能核苷酸，每个核苷酸限定侧链位点和第一单链寡核苷酸单元， 能够特异性结合目标的第一单链多核苷酸序列; 和从所述多功能核苷酸延伸的侧挂多核苷酸侧链，其每个包括能够特异性结合目标的第二单链多核苷酸序列的第二单链寡核苷酸单元的迭代，所有侧链的总迭代次数至少为20。 还提供制备这些多核苷酸的方法。
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2.发明授权Nucleic acid hybridization assays employing large comb-type branched polynucleotides 失效使用大梳状支链多核苷酸的核酸杂交测定
公开(公告)号：US5849481A
公开(公告)日：1998-12-15
申请号：US470124
申请日：1995-06-05
申请人： Michael S. Urdea , Thomas Horn , Chu-An Chang , Brian Warner , Timothy J. Fultz
发明人： Michael S. Urdea , Thomas Horn , Chu-An Chang , Brian Warner , Timothy J. Fultz
摘要： Comb-type branched polynucleotides are used as amplification multimers in nucleic acid hybridization assays.
摘要翻译： 梳状支化多核苷酸用作核酸杂交测定中的扩增多聚体。
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3.发明授权N-4 modified pyrimidine deoxynucleotides and oligonucleotide probes synthesized therewith 失效N-4修饰的嘧啶脱氧核苷酸和与其合成的寡核苷酸探针
公开(公告)号：US5591584A
公开(公告)日：1997-01-07
申请号：US429196
申请日：1995-04-26
申请人： Chu-An Chang , Michael S. Urdea , Thomas Horn
发明人： Chu-An Chang , Michael S. Urdea , Thomas Horn
摘要： Novel oligonucleotide probes are provided having the structure ##STR1## wherein: R.sup.2 is lower alkyl; R.sup.3 is C.sub.1 -C.sub.12 alkylene containing 0 to 6 linkages selected from the group consisting of --O--, --S-- and --NR.sup.12 -- wherein R.sup.12 is hydrogen or lower alkyl; R.sup.5 is hydrogen or lower alkyl; R.sup.6 is selected from the group consisting of hydrogen, methyl, bromo and iodo; R.sup.7 is C.sub.1 -C.sub.12 alkylene containing 0 to 6 linkages selected from the group consisting of --O--, --S-- and --NR.sup.12 --, and bound to R.sup.8 through an --O--, --S-- or --NR.sup.12 -- moiety; and R.sup.8 is a protecting group that can be removed and replaced, without affecting the remainder of the compound, by reduction with a reducing agent. Methods for using the probes are provided as well.
摘要翻译： 提供具有结构“IMAGE”或“IMAGE”的新型寡核苷酸探针，其中：R2是低级烷基; R3是含有0-6个选自-O - ， - S-和-NR12-的键的C1-C12亚烷基，其中R12是氢或低级烷基; R5是氢或低级烷基; R 6选自氢，甲基，溴和碘; R7是含有0-6个选自-O - ， - S-和-NR12-的键的C1-C12亚烷基，并且通过-O - ， - S-或-NR 12 - 部分键合到R8上; 并且R8是可以通过用还原剂还原而可以除去和替代的保护基，而不影响其余的化合物。 还提供了使用探针的方法。
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4.发明授权N-4 modified pyrimidine deoxynucleotides and oligonucleotide probes synthesized therewith 失效N-4修饰的嘧啶脱氧核苷酸和与其合成的寡核苷酸探针
公开(公告)号：US5580731A
公开(公告)日：1996-12-03
申请号：US296793
申请日：1994-08-25
申请人： Chu-An Chang , Michael S. Urdea , Thomas Horn
发明人： Chu-An Chang , Michael S. Urdea , Thomas Horn
摘要： Novel N-4 modified pyrimidine analogs are provided having the structure (I) or (II) ##STR1## wherein: R.sup.1 is selected from the group consisting of hydrogen, acid-sensitive, base-stable blocking groups and acyl capping groups; R.sup.2 is lower alkyl; R.sup.3 is C.sub.1 -C.sub.12 alkylene containing 0 to 6 linkages selected from the group consisting of --0--, --S-- and --NH--; R.sup.4 is ##STR2## in which R.sup.9 is hydrogen or an optionally substituted aliphatic group; R.sup.10 and R.sup.11 are hydrocarbyl or together form a mono- or polyheterocyclic ring; R.sup.5 is hydrogen or lower alkyl; R.sup.6 is selected from the group consisting of hydrogen, methyl, bromo and iodo; R.sup.7 is C.sub.1 -C.sub.12 alkylene containing 0 to 6 linkages selected from the group consisting of --O--, --S-- and --NR.sup.12 -- wherein R.sup.12 is hydrogen or lower alkyl; and R.sup.8 is a protecting group that can be removed and replaced by reduction. Synthetic methods for preparing the compounds are provided as well, as are methods for making and using polynucleotide probes containing the novel pyrimidine analogs.
摘要翻译： 提供具有结构（I）或（II）的新型N-4修饰的嘧啶类似物，其中：R1选自氢，酸敏感，碱稳定 封闭基团和酰基封端基团; R2是低级烷基; R3是含有0-6个选自-O - ， - S - 和-NH-的键的C1-C12亚烷基; R4是
，其中R9是氢或任选取代的脂族基; R 10和R 11是烃基或一起形成单或多杂环; R5是氢或低级烷基; R 6选自氢，甲基，溴和碘; R7是含有0-6个选自-O - ， - S-和-NR12-的键的C1-C12亚烷基，其中R12是氢或低级烷基; R8是可以除去并被还原代替的保护基。 还提供了制备化合物的合成方法，以及制备和使用含有新型嘧啶类似物的多核苷酸探针的方法。 -
公开(公告)号：US5747244A
公开(公告)日：1998-05-05
申请号：US813338
申请日：1991-12-23
IPC分类号： C12N15/09 , C07H21/04 , C12Q1/68 , C12Q1/70 , C12R1/92 , G01N33/53 , G01N33/547 , G01N33/566 , C07K17/08 , C08L89/00
CPC分类号： C12Q1/6813 , C12Q1/6834
摘要： Nucleic acid probes are immobilized on polystyrene surfaces such as the wells of microtiter plates for use in solution phase nucleic acid sandwich hybridization assays, particularly those using large branched DNA amplification multimers, by: (a) cleansing the surface by washing it sequentially with a strong acid, a strong base, and water, (b) passively adsorbing a polypeptide having primary amino groups onto the cleansed surface, and (c) covalently bonding the probe to the adsorbed polypeptide via a base-stable bifunctional crosslinking agent, and (d) subjecting the surface to conditions that simulate the hybridization conditions used in the assay.
摘要翻译： 将核酸探针固定在聚苯乙烯表面上，例如微量滴定板的孔，用于溶液相核酸夹心杂交测定法，特别是使用大分支DNA扩增多聚体的那些;（a）通过以下方式顺序洗涤来清洗表面： 酸，强碱和水，（b）被动吸附具有伯氨基的多肽到清洁表面上，和（c）通过碱稳定的双功能交联剂将探针共价结合到吸附的多肽，和（d） 使表面经受模拟测定中使用的杂交条件的条件。
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公开(公告)号：US5712383A
公开(公告)日：1998-01-27
申请号：US438639
申请日：1995-05-10
IPC分类号： C12N15/09 , C07H21/04 , C12Q1/68 , C12Q1/70 , C12R1/92 , G01N33/53 , G01N33/547 , G01N33/566 , C07H21/02
CPC分类号： C12Q1/6813 , C12Q1/6834
摘要： Nucleic acid probes are immobilized on polystyrene surfaces such as the wells of microtiter plates for use in solution phase nucleic acid sandwich hybridization assays, particularly those using large branched DNA amplification multimers, by: (a) cleansing the surface by washing it sequentially with a strong acid, a strong base, and water, (b) passively adsorbing a polypeptide having primary amino groups onto the cleansed surface, and (c) covalently bonding the probe to the adsorbed polypeptide via a base-stable bifunctional crosslinking agent, and (d) subjecting the surface to conditions that simulate the hybridization conditions used in the assay.
摘要翻译： 将核酸探针固定在聚苯乙烯表面上，例如微量滴定板的孔，用于溶液相核酸夹心杂交测定法，特别是使用大分支DNA扩增多聚体的那些;（a）通过以下方式顺序洗涤来清洗表面： 酸，强碱和水，（b）被动吸附具有伯氨基的多肽到清洁表面上，和（c）通过碱稳定的双功能交联剂将探针共价结合到吸附的多肽，和（d） 使表面经受模拟测定中使用的杂交条件的条件。
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公开(公告)号：US4914210A
公开(公告)日：1990-04-03
申请号：US104200
申请日：1987-10-02
申请人： Corey Levenson , Chu-An Chang , Fred T. Oakes
发明人： Corey Levenson , Chu-An Chang , Fred T. Oakes
IPC分类号： C07C43/11 , C07C43/178 , C07C323/12 , C07D209/48 , C07D317/22 , C07F9/24 , C07F9/26 , C07F9/48 , C07F9/553 , C07H21/00 , C07H21/04 , C08G65/335 , C12Q1/68 , G01N33/535
CPC分类号： C07D209/48 , C07C323/12 , C07C43/11 , C07C43/178 , C07D317/22 , C07F9/2408 , C07F9/2441 , C07F9/2458 , C07F9/2487 , C07F9/26 , C07F9/5537 , C07H21/00 , C08G65/3356 , C12Q1/6816 , G01N33/535
摘要： Oligonucleotide functionalizing reagents are disclosed which are useful in introducing sulfhydryl, amino and additional hydroxyl groups into oligonucleotides. The reagents are substantially linear in structure, at one end provided with a phosphoramidite moiety, at an opposing end provided with a sulfhydryl, amino or hydroxyl moiety, the two ends linked through a hydrophilic spacer chain. Methods of using and synthesizing the novel reagents are disclosed as well.
摘要翻译： 公开了寡核苷酸官能化试剂，其用于将巯基，氨基和另外的羟基引入寡核苷酸。 试剂在结构上基本上是线性的，在一端具有亚磷酰胺部分，在相对端具有巯基，氨基或羟基部分，两个末端通过亲水间隔链连接。 还公开了使用和合成新试剂的方法。
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公开(公告)号：US07534569B2
公开(公告)日：2009-05-19
申请号：US11770787
申请日：2007-06-29
申请人： Chu-An Chang , Shiaw-Min Chen
发明人： Chu-An Chang , Shiaw-Min Chen
CPC分类号： C12N15/1031
摘要： A method of generating a single-stranded nucleic acid molecule comprising (a) combining in a mixture under conditions suitable for a polymerase extension reaction, (i) a polymerase, (ii) an initial polynucleotide comprising a 5′ portion and a 3′ portion, wherein the polynucleotide forms the nucleic acid molecule 5′ end; and (iii) a plurality of overlapping template oligonucleotides each having a 5′ template portion and a 3′ portion. The method further comprises (b) hybridizing the initial polynucleotide or the extension polynucleotide and one of the template oligonucleotides; (c) incubating the mixture for sufficient time to allow an extension polynucleotide to be synthesized; (d) adding a competimer that competes with the template oligonucleotide in step (b); (e) denaturing the extension polynucleotide and template oligonucleotide; and (f) repeating steps (b), (c), (d), and (e) to generate the single-stranded nucleic acid molecule, wherein the number of repeated cycles equals the number of different template oligonucleotides.
摘要翻译： 一种产生单链核酸分子的方法，包括（a）在适于聚合酶延伸反应的条件下在混合物中混合，（i）聚合酶，（ii）包含5'部分和3'部分的初始多核苷酸 ，其中所述多核苷酸形成核酸分子5'末端; 和（iii）多个具有5'模板部分和3'部分的重叠模板寡核苷酸。 该方法还包括（b）使初始多核苷酸或延伸多核苷酸与模板寡核苷酸之一杂交; （c）将混合物温育足够的时间以允许合成延伸的多核苷酸; （d）在步骤（b）中加入与模板寡核苷酸竞争的竞争体; （e）使延伸多核苷酸和模板寡核苷酸变性; 和（f）重复步骤（b），（c），（d）和（e）以产生单链核酸分子，其中重复循环的数量等于不同模板寡核苷酸的数目。
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公开(公告)号：US4859419A
公开(公告)日：1989-08-22
申请号：US19923
申请日：1987-02-27
CPC分类号： B01L3/5025 , G01N21/253 , G01N33/5302 , G01N33/5304 , G01N21/29 , G01N21/78 , G01N35/02 , Y10S435/805 , Y10S435/97 , Y10S435/974
摘要： Apparatus for immunoassay of multiple samples of biological fluids (e.g., AIDS antibodies), the apparatus being capable of mounting for rocking during incubation. A frame receives plural test vessels so that the vessels are interconnected for circulation through all the vessels of buffers, de-ionized water and air and for discharge of waste. Each vessel is transparent and contains nitrocellulose test paper impregnated with antigens to the antibodies. Valved ports in each vessel accept sequential insertion of the biological fluids, an enzyme conjugate and color developer. Color changes in the individual test papers are observed through the transparent vessels and identify presence of antibodies in the samples.
摘要翻译： 用于免疫测定多种生物流体样品（例如AIDS抗体）的装置，该装置能够在孵育期间安装摇摆。 框架接收多个测试容器，使得容器互连以便通过缓冲器，去离子水和空气的所有容器循环并排出废物。 每个容器是透明的，并含有用抗原浸渍抗体的硝化纤维素试纸。 每个容器中的阀门接受生物流体，酶缀合物和显色剂的顺序插入。 通过透明容器观察单个测试纸中的颜色变化，并鉴定样品中抗体的存在。
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