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公开(公告)号：US5422271A
公开(公告)日：1995-06-06
申请号：US979569
申请日：1992-11-20
CPC分类号： B01L3/502
摘要： A device and method are disclosed for amplifying and detecting nucleic acid material. The device and method use a label and signalling material responsive to the label to produce a detectable signal. A surprising result of the method and device is that at least one of the wash steps heretofore required has been eliminated without substantially adversely affecting the results.
摘要翻译： 公开了用于扩增和检测核酸材料的装置和方法。 该装置和方法使用响应标签的标签和信令材料产生可检测信号。 该方法和装置的令人惊奇的结果是至少需要一个洗涤步骤已被消除，而不会对结果产生实质性的不利影响。
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2.发明授权Diagnostic compositions, elements, methods and test kits for amplification and detection of retroviral DNA using primers having matched melting temperatures 失效使用具有匹配熔融温度的引物扩增和检测逆转录病毒DNA的诊断组合物，元件，方法和测试试剂盒
公开(公告)号：US5403707A
公开(公告)日：1995-04-04
申请号：US062022
申请日：1993-05-14
CPC分类号： C12Q1/686 , C12Q1/6834
摘要： An aqueous composition containing primers for opposing strands of a target retroviral DNA (such as HIV-I DNA) can be used in polymerase chain reaction to provide simultaneously rapid and efficient amplification and detection of that target DNA and one or more additional target DNA's. The primers for each target DNA differ in length by no more than 5 nucleotides and have a T.sub.m within the range of from about 65.degree. to about 74.degree. C., while all of the T.sub.m 's are within about 5.degree. C. of each other. Such compositions are useful in diagnostic test kits and methods for amplification and detection of multiple nucleic acids, or in "multiplexing", using multiple capture probes, all of which have T.sub.m 's which are greater than 50.degree. C. and within 15.degree. C. of each other.
摘要翻译： 可以在聚合酶链反应中使用含有针对靶向逆转录病毒DNA（例如HIV-1 DNA）的相对链的引物的含水组合物，以便同时快速且有效地扩增和检测该靶DNA和一种或多种另外的靶DNA。 每个靶DNA的引物长度不等于5个核苷酸，Tm在约65-74℃范围内，而所有Tm在彼此的约5℃内。 这样的组合物可用于诊断测试试剂盒和用于多重核酸的扩增和检测的方法，或使用多个捕获探针进行“多路复用”，所有这些探针具有大于50℃的Tm和15℃以内的Tm 彼此。
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3.发明授权Diagnostic and amplification methods using primers having thymine at 3' end to overcome primer-target mismatch at the 3' end 失效在3'末端使用3'末端终止重载目标错误的引导符的诊断和放大方法
公开(公告)号：US5196305A
公开(公告)日：1993-03-23
申请号：US406221
申请日：1989-09-12
申请人： John B. Findlay , Lynn Bergmeyer
发明人： John B. Findlay , Lynn Bergmeyer
CPC分类号： C12Q1/6858 , C12Q1/703 , Y10S435/805 , Y10S435/948 , Y10S436/811
摘要： Methods for amplifying and detecting a predetermined target nucleic acid in a biological specimen are accomplished even where there is a mismatch in a single position between a primer and the target nucleic acid. The mismatch is located at or near the 3' end of the primer. Such a mismatch is overcome using a primer having a nucleotide with a thymine base at the position of the mismatch. The use of such primers is most likely to prime the target and form primer extension products. This method is particularly useful for detection of a nucleic acid sequence which is not fully known, or where there is considerable heterogeneity in DNA target from patient samples.
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4.发明授权Specific binding analytical and separation methods using carboxy containing polymers 失效使用含羧基聚合物的特异性结合分析和分离方法
公开(公告)号：US5262297A
公开(公告)日：1993-11-16
申请号：US876672
申请日：1992-04-30
申请人： Richard C. Sutton , Susan J. Danielson , John B. Findlay , Fred. T. Oakes , Marsha D. B. Oenick , Ignazio S. Ponticello , Harold C. Warren, III
发明人： Richard C. Sutton , Susan J. Danielson , John B. Findlay , Fred. T. Oakes , Marsha D. B. Oenick , Ignazio S. Ponticello , Harold C. Warren, III
IPC分类号： C12Q1/68 , C12Q1/70 , G01N33/543 , G01N33/545 , G01N83/545 , G01N33/546 , G01N33/547
CPC分类号： G01N33/54313 , C12Q1/6876 , C12Q1/701 , C12Q1/703 , G01N33/545 , Y10S436/804 , Y10S436/814 , Y10S436/817 , Y10S436/818 , Y10S436/824
摘要： Specific binding methods are used for diagnostic assays and purification separations whereby the specific binding capture reagent is prepared from copolymers having highly reactive carboxy groups. These groups are extended from the polymer surface with a linking group having from 8 to 50 atoms in the chain and two or more alkylene, arylene, alkylenearylene or arylenealkylene groups. To these reactive groups is attached a biologically active substance such as a protein or oligonucleotide which then participates in the diagnostic assays or purification separation methods.
摘要翻译： 特异性结合方法用于诊断测定和纯化分离，由此特异性结合捕获试剂由具有高反应性羧基的共聚物制备。 这些基团从聚合物表面延伸，在链中具有8至50个原子的连接基团和两个或更多个亚烷基，亚芳基，亚烷基亚芳基或亚芳基亚烷基。 将这些反应性基团连接到生物活性物质如蛋白质或寡核苷酸上，然后参与诊断测定或纯化分离方法。
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公开(公告)号：US5229297A
公开(公告)日：1993-07-20
申请号：US962159
申请日：1992-10-15
申请人： Paul N. Schnipelsky , Leonard J. Seaberg , Charles C. Hinckley , Jeffrey A. Wellman , William H. Donish , John B. Findlay
发明人： Paul N. Schnipelsky , Leonard J. Seaberg , Charles C. Hinckley , Jeffrey A. Wellman , William H. Donish , John B. Findlay
IPC分类号： B01L3/00 , C12Q1/68 , G01N33/543 , G01N35/00
CPC分类号： G01N33/54313 , B01L3/502 , B01L2300/0636 , B01L2300/0816 , B01L2300/0867 , B01L2400/0478 , B01L2400/0481 , G01N33/54326 , G01N35/0098 , Y10T436/143333 , Y10T436/2575
摘要： A cuvette and a method of use which prevent nucleic acid amplified by PCR technology from being released to the atmosphere, while still proceeding to a detection step to determine whether or not the nucleic acid is present. Detection reagents are either pre-incorporated into compartments in the cuvette or added after amplification. In the latter case, a check valve prevents amplified nucleic acid from being released. Transfer of liquids between compartments is achieved via the use of flexible compartment walls and an external pressure source, or via pistons that are part of the cuvette and operate on the compartments as a piston within a piston chamber.
摘要翻译： 将通过PCR技术扩增的核酸防止释放到大气中的试管和使用方法，同时仍然进行检测步骤以确定核酸是否存在。 检测试剂预先掺入比色皿的隔室或扩增后加入。 在后一种情况下，止回阀防止扩增的核酸被释放。 通过使用柔性隔室壁和外部压力源，或通过作为反应杯的一部分的活塞，并且在作为活塞室内的活塞的隔室上操作来实现隔室之间的液体传递。
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6.发明授权Methods for capture and selective release of nucleic acids using weakly basic polymer and amplification of same 失效使用弱碱性聚合物捕获和选择性释放核酸的方法及其扩增
公开(公告)号：US5582988A
公开(公告)日：1996-12-10
申请号：US306870
申请日：1994-09-15
申请人： John W. Backus , Tobias E. Ekeze , Jerome C. Swartz , Richard C. Sutton , Ignazio S. Ponticello , JoAnne H. Kerschner , John B. Findlay
发明人： John W. Backus , Tobias E. Ekeze , Jerome C. Swartz , Richard C. Sutton , Ignazio S. Ponticello , JoAnne H. Kerschner , John B. Findlay
IPC分类号： C12N15/09 , C07H21/04 , C07K14/045 , C07K14/16 , C07K14/35 , C08F20/34 , C08F20/60 , C12N15/10 , C12P19/34 , C12Q1/68 , C12Q1/70 , G01N33/543 , G01N33/569
CPC分类号： C12N15/1003 , C07K14/005 , C07K14/35 , C08F20/34 , C08F20/60 , C12N15/1006 , C12Q1/6806 , C12N2710/16122 , C12N2740/16022
摘要： Nucleic acids can be made available for amplification or other treatment after lysis by contacting the lysate with specific weakly basic polymers to form a precipitate with the nucleic acids at acidic pH. After removing non-precipitated materials, the pH is then made basic, thereby releasing the nucleic acids from the polymer. This method for preparing specimen samples is simple and quite rapid, and the released nucleic acids can be further treated in hybridization assays or amplification procedures. The weakly basic polymers are water-soluble and cationic at acidic pH, but neutral in charge at basic pH.
摘要翻译： 通过使溶解产物与特定的弱碱性聚合物接触以在酸性pH下与核酸形成沉淀物，使核酸可用于扩增或裂解后的其它处理。 除去非沉淀物后，将pH调至碱性，从而将聚合物中的核酸释放出来。 制备标本样品的这种方法是简单且相当快速的，并且释放的核酸可以在杂交测定或扩增程序中进一步处理。 弱碱性聚合物在酸性pH下为水溶性和阳离子型，而在碱性pH下为负电荷。
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公开(公告)号：US5328825A
公开(公告)日：1994-07-12
申请号：US847447
申请日：1992-03-06
IPC分类号： C12N15/09 , C12Q1/68 , C12Q1/70 , G01N33/543 , C07H21/04
CPC分类号： G01N33/54313 , C12Q1/6834 , C12Q1/703
摘要： An oligonucleotide is linked to a particle through a protein or carbohydrate to form a water-insoluble nucleic acid probe. The protein or carbohydrate has a pI of about 6 or less, and has been chemically modified with an acylating, alkylating or sulfonylating agent. The particle surface is substantially free of other proteins or carbohydrates. The probe is useful in various diagnostic and purification methods wherein hybridization of the oligonucleotide with a target nucleic acid is possible. In one instance, the probe can be used to capture a DNA strand which has been amplified using polymerase chain reaction techniques.
摘要翻译： 寡核苷酸通过蛋白质或碳水化合物与颗粒连接以形成水不溶性核酸探针。 蛋白质或碳水化合物具有约6或更小的pI，并已用酰化，烷基化或磺酰化剂进行化学修饰。 颗粒表面基本上不含其他蛋白质或碳水化合物。 探针可用于各种诊断和纯化方法，其中寡核苷酸与靶核酸的杂交是可能的。 在一个实例中，探针可用于捕获已经使用聚合酶链反应技术扩增的DNA链。
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8.发明授权Biologically active reagents prepared from carboxy-containing polymer, analytical element and methods of use 失效含羧基聚合物制备的生物活性试剂，分析元素和使用方法
公开(公告)号：US5147777A
公开(公告)日：1992-09-15
申请号：US539774
申请日：1990-06-18
申请人： Richard C. Sutton , Susan J. Danielson , John B. Findlay , Fred T. Oakes , Marsha D. B. Oenick , Ignazio S. Ponticello , Harold C. Warren, III
发明人： Richard C. Sutton , Susan J. Danielson , John B. Findlay , Fred T. Oakes , Marsha D. B. Oenick , Ignazio S. Ponticello , Harold C. Warren, III
IPC分类号： C08F8/00 , C08F246/00 , C12Q1/68 , C12Q1/70 , G01N33/53 , G01N33/543 , G01N33/545
CPC分类号： G01N33/54313 , C12Q1/6876 , C12Q1/701 , C12Q1/703 , G01N33/545 , Y10S436/805 , Y10T428/2991 , Y10T428/2998
摘要： Biologically active reagents are prepared from particles of copolymers having highly reactive carboxy or equivalent groups. The reagents are prepared by covalently attaching biologically active substances, for example antibodies, to the particles, directly or indirectly through highly reactive carboxy groups on the particle surface. These reagents are used to advantage in analytical elements, methods for the detection of specific binding ligands (such as immunological species) and immunoassays, and in purification methods as affinity chromatography reagents.
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9.发明授权Analytical element containing photosensitive compound and filter layer and method of use 失效含有光敏化合物和过滤层的分析元素及使用方法
公开(公告)号：US4788140A
公开(公告)日：1988-11-29
申请号：US830036
申请日：1986-02-18
CPC分类号： G01N33/525 , C12Q1/50 , Y10S435/805
摘要： A multilayer analytical element for the determination of a clinically significant enzyme analyte comprises a photosensitive compound (e.g. a photosensitive dye or dye precursor) and a filter layer containing one or more filter dyes. The filter layer is situated in the element such that incident radiation used to detect a density change resulting from interaction of the analyte and the photosensitive compound passes through the filter layer before it reaches the photosensitive compound. The use of the filter layer inhibits premature changes in the photosensitive compound caused by incident radiation. This element is particularly useful for the determination of creatine kinase or one of its isoenzymes, e.g. creatine kinase-MB.
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10.发明授权Composition, analytical element and method for the quantification of creatine kinase 失效组成，分析元素和定量肌酸激酶的方法
公开(公告)号：US4547461A
公开(公告)日：1985-10-15
申请号：US459026
申请日：1983-01-18
CPC分类号： C12Q1/48 , C12Q1/28 , C12Q1/50 , Y10S435/805 , Y10S435/81
摘要： An enzymatic method for the analytical determination of creatine kinase in an aqueous liquid such as blood serum is described. The determination is made by measuring an optical density change using the reagents creatine phosphate, adenosine diphosphate, glycerol, glycerol kinase, .alpha.-glycerophosphate oxidase, a chromagen and a mercapto-containing creatine kinase activator. The mercapto-containing creatine kinase activator is added in encapsulated form or in low concentrations so as to preserve the activity of the chromagen.
摘要翻译： 描述了在诸如血清的水性液体中分析肌酸激酶的分析测定方法。 通过使用试剂磷酸肌酸，二磷酸腺苷，甘油，甘油激酶，α-甘油磷酸酯氧化酶，色素和含巯基的肌酸激酶激活剂测量光密度变化来进行测定。 含巯基的肌酸激酶活化剂以胶囊形式或低浓度加入，以保持色素的活性。
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