公开(公告)号：US06635439B2

公开(公告)日：2003-10-21

申请号：US09745212

申请日：2000-12-21

申请人： Todd Morrison ,  Veronica V. Thralls ,  Franklin A. Jurik

发明人： Todd Morrison ,  Veronica V. Thralls ,  Franklin A. Jurik

IPC分类号： C12Q128

CPC分类号： G01N31/228 ,  A61L2/28 ,  C12Q1/22 ,  C12Q1/28 ,  G01N21/8483

摘要： A method for determining exposure of a test sample to hydrogen peroxide in an atmosphere includes placing a substrate into the atmosphere, contacting the substrate with the hydrogen peroxide, reacting the hydrogen peroxide with a dye intermediate to produce a chromophore indicative of the integrated exposure of hydrogen peroxide in the atmosphere; and correlating the chromophore to the integrated exposure of hydrogen peroxide in the atmosphere. The dye intermediate may be a dye-couple of 3-methyl-2-benzothiazolinone hydrazone hydrochloride (MBTH) and 3-dimethylaminobenzoic acid (DMAB). The intensity of the chromophore is indicative of an integrated hydrogen peroxide exposure.

公开(公告)号：US06503726B2

公开(公告)日：2003-01-07

申请号：US09794789

申请日：2001-02-27

申请人： Lakshmi Anne ,  Weixing Luo ,  Ker-kong Tung

发明人： Lakshmi Anne ,  Weixing Luo ,  Ker-kong Tung

IPC分类号： C12Q128

CPC分类号： C12Q1/28 ,  Y10S435/97 ,  Y10T436/173845

摘要： A device and method to detect oxidant adulterants added to a sample to prevent detection of tetrahydrocannabinol and/or tetrahydrocannabinol metabolites. The device and method detect popularly available adulterants such as chromate and nitrite through their reaction with a chromogenic substrate for a peroxidase enzyme. The substrate may be applied to a pad. A change in color in the presence of the oxidant may be detected directly or indirectly. The device and method may be used to detect the presence of any oxidant adulterant and is not specific for only one or a few adulterants added to a urine sample submitted for drug testing.

公开(公告)号：US06617125B2

公开(公告)日：2003-09-09

申请号：US09895937

申请日：2001-06-29

申请人： Karl Edwin Adler, Jr.

发明人： Karl Edwin Adler, Jr.

IPC分类号： C12Q128

CPC分类号： C12N9/96 ,  C12Q1/28 ,  Y10S435/968 ,  Y10S435/975

摘要： A method for enhancing the conversion of a phenol substrate to a product by a peroxidase enzyme comprises the steps of reacting a conjugate comprising a detectably labeled phenol-containing molecule with a peroxidase enzyme in the presence of an enhancing reagent, the enhancing reagent comprising an organic compound having the structure wherein each R is independently selected from the group consisting of: hydrogen and a C1-12 substituent; where V, W, Y and Z are each independently selected from the group consisting of: H, halogen, the C1-12 substituent, NR2, OR and SR; and where X is selected from the group consisting of: H, Br, Cl, F, the C1-12 substituent, NR2, OR and SR; or synergistic mixtures of an inorganic salt and the organic compound. The C1-12 substituent is linear, branched or cyclic. The C1-12 substituent is alkyl, alkenyl, alkynyl, heteroatom substituted alkyl, heteroatom substituted alkenyl, heteroatom substituted alkynyl, aryl, arylalkyl, arylalkenyl or arylalkynyl. Further, the heteroatom is N, O, S or halogen. Any C, N, O or S in the C1-12 substituent optionally has a pendant moiety which is carbonyl, hydroxyl, carboxyl, amine, thiol, thioester, thioether, phosphate, alkoxy, aryl, arylalkyl, sulfonamide or alkyl halide. A kit is provided containing an enhancing reagent for enhancing the detection of an enzyme reaction as described herein, together with instructions for use.

摘要翻译： 通过过氧化物酶增强酚底物转化成产物的方法包括以下步骤：在增强剂存在下使包含可检测标记的含酚分子的结合物与过氧化物酶反应，所述增强剂包含有机 具有各R的结构的化合物独立地选自：氢和C1-12取代基; 其中V，W，Y和Z各自独立地选自：H，卤素，C 1-12取代基，NR 2，OR和SR; 并且其中X选自：H，Br，Cl，F，C 1-12取代基，NR 2，OR和SR; 或无机盐和有机化合物的协同混合物。 C1-12取代基是直链，支链或环状的。 C1-12取代基是烷基，烯基，炔基，杂原子取代的烷基，杂原子取代的烯基，杂原子取代的炔基，芳基，芳基烷基，芳基烯基或芳基炔基。 此外，杂原子是N，O，S或卤素。 C1-12取代基中任何C，N，O或S任选具有羰基，羟基，羧基，胺，硫醇，硫酯，硫醚，磷酸酯，烷氧基，芳基，芳基烷基，磺酰胺或烷基卤的侧基部分。 提供了一种试剂盒，其含有用于增强本文所述的酶反应的检测的增强剂以及使用说明书。

公开(公告)号：US06432662B1

公开(公告)日：2002-08-13

申请号：US09171334

申请日：1999-05-20

申请人： Paul D. Davis ,  Kelli D. Feather-Henigan ,  Kimberly Hines

发明人： Paul D. Davis ,  Kelli D. Feather-Henigan ,  Kimberly Hines

IPC分类号： C12Q128

CPC分类号： C12Q1/28

摘要： Chemiluminescent detection of molecules of synthetic or natural origin such as proteins and nucleic acids (DNA and RNA), as well as other biologic molecules, is increasingly replacing radioactive detection as the method of choice where sensitivity is critical. In such assays, luminescence is customarily achieved by the oxidation of a luminol or isoluminol substrate in the presence of an oxidizing agent such as hydrogen peroxide or hydrogen peroxide source, such as perborate, and a peroxidase catalyst such as horseradish peroxidase. To obtain useful levels of luminescence (e.g., detectable levels) by customary techniques, a luminescent enhancer is also present during oxidation. It has been found in the practice of the present invention that azine enhancers have contained an impurity which reduces the properties of the chemiluminescent assay working solutions. The present invention describes a working solution, a process of using the working solution, and kits containing solutions which can be mixed to form the working solutions comprising a solution which is useful for the chemiluminescent assay of peroxidase activity comprising: a) at least one chemiluminescent cyclic diacylhydrazide, b) at least one azine enhancer, and c) at least one oxidizine agent wherein said azine enhancer comprises less than 0.005 parts mole/mole total basis of azine compounds having a hydrogen atom bonded to a nitrogen atom of the azine ring. It has generally been found that this hydrogen containing azine is a poisoning agent for the performance of the azine enhancer.

摘要翻译： 化学发光检测合成或天然来源的分子，如蛋白质和核酸（DNA和RNA）以及其他生物分子，正在越来越多地将放射性检测替代为灵敏度至关重要的选择方法。 在这种测定中，通常在氧化剂如过氧化氢或过氧化氢源（例如过硼酸盐）和过氧化物酶催化剂如辣根过氧化物酶的存在下氧化鲁米诺或异鲁米酚底物来实现发光。 为了通过常规技术获得有用的发光水平（例如可检测水平），在氧化期间也存在发光增强剂。 在本发明的实践中已经发现，吖嗪增强剂含有降低化学发光测定工作溶液性质的杂质。 本发明描述了工作解决方案，使用工作溶液的方法以及包含可混合以形成包含可用于过氧化物酶活性的化学发光测定的溶液的工作溶液的试剂盒，所述方法包括：a）至少一种化学发光 环二酰基酰肼，b）至少一种吖嗪增效剂，和c）至少一种氧化剂，其中所述吖嗪增强剂包含具有与吖嗪环的氮原子键合的氢原子的吖嗪化合物的总计基于0.005摩尔/摩尔。 通常已经发现，该含氢吖嗪是用于实施吖嗪增强剂的中毒剂。

公开(公告)号：US06265179B1

公开(公告)日：2001-07-24

申请号：US09495882

申请日：2000-02-01

申请人： Mingjie Zhou ,  Richard P. Haugland

发明人： Mingjie Zhou ,  Richard P. Haugland

IPC分类号： C12Q128

CPC分类号： C12Q1/26 ,  C12Q1/42 ,  C12Q1/48 ,  G01N2333/91102 ,  G01N2333/91142 ,  G01N2333/916 ,  Y10S435/968 ,  Y10S435/975

摘要： Inorganic phosphate may be detected and optionally quantified via the coupling of a phosphate-dependent enzymatic reaction with an enzyme system that generates hydrogen peroxide in the presence of a chromogenic or fluorogenic peroxidase substrate. Phosphate consuming or phosphate-producing enzymes or their substrates may also be detected and/or quantified, including pyrophosphatase enzymes or pyrophosphatase.

摘要翻译： 可以通过磷酸盐依赖性酶反应与在显色或荧光过氧化物酶底物存在下产生过氧化氢的酶系统的偶联来检测和任选地定量无机磷酸盐。 磷酸盐消耗或磷酸盐生成酶或其底物也可被检测和/或定量，包括焦磷酸酶或焦磷酸酶。

公开(公告)号：US06602679B2

公开(公告)日：2003-08-05

申请号：US09770592

申请日：2001-01-26

申请人： Brij Giri

发明人： Brij Giri

IPC分类号： C12Q128

CPC分类号： C12Q1/28 ,  C12Q1/66 ,  G01N33/52

摘要： A substrate having an extended shelf life for chemiluminescent detection and assaying is useful with enzyme-based probes as well as for immunoassays, DNA or RNA or protein detection and blotting. The substrate is a mixture of (a) a chemiluminescent compound, (b) an oxidant, (c) a stabilizer and (d) a buffer. The substrate may further include a chemiluminescent enhancer and a solubilizer. The substrate may be in an organic solvent or may be aqueous based.

摘要翻译： 具有延长的化学发光检测和测定保质期的底物可用于酶基探针以及免疫测定，DNA或RNA或蛋白质检测和印迹。 底物是（a）化学发光化合物，（b）氧化剂，（c）稳定剂和（d）缓冲剂的混合物。 底物还可以包括化学发光增强剂和增溶剂。 底物可以是有机溶剂，也可以是水性的。

公开(公告)号：US06599713B1

公开(公告)日：2003-07-29

申请号：US09913161

申请日：2001-08-10

申请人： Yoshihiro Hatanaka ,  Ryo Serizawa

发明人： Yoshihiro Hatanaka ,  Ryo Serizawa

IPC分类号： C12Q128

CPC分类号： C12Q1/28 ,  G01N33/5094 ,  G01N33/54393 ,  G01N33/56972 ,  G01N33/573 ,  G01N2333/902 ,  G01N2333/908

摘要： Disclosed is a method for determining a white blood cell count of a whole blood sample, which comprises: mixing a whole blood sample with a surfactant to thereby obtain a mixture; allowing the mixture to stand for a time sufficient to lyze the white blood cells contained in the whole blood sample and release intrinsic myeloperoxidase from the white blood cells; measuring the concentration of the released myeloperoxidase in the mixture; and determining the white blood cell count in the whole blood sample, based on the concentration of the released myeloperoxidase. Also disclosed is a method of the present invention, in which, in addition to a white blood cell count of a whole blood sample, the concentration of C-reactive protein contained in the same whole blood sample is measured.

摘要翻译： 公开了一种测定全血样品的白细胞计数的方法，其特征在于：将全血样品与表面活性剂混合，得到混合物; 使混合物静置足以裂解全血样品中所含的白细胞并从白细胞中释放本征髓过氧化物酶; 测量混合物中释放的髓过氧化物酶的浓度; 并根据所释放的髓过氧化物酶的浓度确定全血样品中的白细胞计数。 还公开了本发明的方法，其中，除了全血样品的白细胞计数之外，还测定同一全血样品中所含的C-反应蛋白的浓度。

公开(公告)号：US06221624B1

公开(公告)日：2001-04-24

申请号：US09445802

申请日：2000-02-09

申请人： Allan Otto Fog Lihme ,  Margit Wikborg

发明人： Allan Otto Fog Lihme ,  Margit Wikborg

IPC分类号： C12Q128

CPC分类号： G01N33/581 ,  G01N33/535 ,  Y10S435/968

摘要： The present invention relates to a method of using a storage-stable, pre-stained 3,3′,5,5′-tetramethylbenzidine (TMB) substrate in an enzyme substrate. The TMB substrates comprise: (a) a water-based TMB substrate composition which includes a peroxide, e.g., hydrogen peroxide or urea hydrogen peroxide, and (b) a dye which is soluble therein. Suitable dyes to be included in the substrate preferably have an absorbance at 450 nm of at the most milli absorbance units. Examples of dues are Pholixine B, Eosin B and Quinaldine Red. The substrates are advantageous in that they include a dye which is visible to the human eye. This is especially valuable when the substrates are handled and measured. The substrates are especially suitable for enzyme assays such as enzyme-linked-immunosorbent-assays (ELISA), e.g. horse radish peroxide (HRP) is used. The substrates may advantageously include less than 5% organic solvents and a solubility increasing agent such as polyvinylalcohol. The substrates are storage-stable for more than 12 months.

摘要翻译： 本发明涉及在酶底物中使用储存稳定的预染色的3,3'，5,5'-四甲基联苯胺（TMB）底物的方法。 TMB底物包括：（a）水性TMB底物组合物，其包含过氧化物，例如过氧化氢或尿素过氧化氢，和（b）可溶于其中的染料。 包含在底物中的合适的染料优选在450nm处具有最大吸光度单位的吸光度。 膳食的例子有Pholixine B，Eosin B和Quinaldine Red。 这些基材的优点在于它们包括人眼可见的染料。 当处理和测量基板时，这是特别有价值的。 底物特别适合于酶测定，例如酶联免疫吸附测定（ELISA），例如酶联免疫吸附测定（ELISA）。 使用过氧化萝卜（HRP）。 底物可以有利地包括小于5％的有机溶剂和增溶剂如聚乙烯醇。 底物储存稳定超过12个月。

公开(公告)号：US06767717B1

公开(公告)日：2004-07-27

申请号：US09914268

申请日：2002-02-15

申请人： Harel Itzhaky ,  Ehud Keinan

发明人： Harel Itzhaky ,  Ehud Keinan

IPC分类号： C12Q128

CPC分类号： C12Q1/28 ,  Y10S435/81 ,  Y10T436/200833 ,  Y10T436/206664

摘要： The present invention provides a method of detecting a peroxide-based explosive in a sample suspected of consisting of or comprising such explosive, which method comprises dissolving said sample in a suitable organic solvent, contacting the solution with an aqueous solution of a strong acid capable of decomposing said explosive to release hydrogen peroxide, and contacting the resulting mixture with a peroxidase enzyme. The invention also provides a kit for use in the method of the invention.

摘要翻译： 本发明提供了一种检测疑似由或包含该炸药的样品中的过氧化物类炸药的方法，该方法包括将所述样品溶解在合适的有机溶剂中，使该溶液与强酸的水溶液接触， 分解所述炸药以释放过氧化氢，并使所得混合物与过氧化物酶接触。 本发明还提供了用于本发明方法的试剂盒。

公开(公告)号：US06593100B2

公开(公告)日：2003-07-15

申请号：US09859988

申请日：2001-05-17

申请人： Mark Norman Bobrow ,  Karl Edwin Adler ,  Kevin Aaron Roth

发明人： Mark Norman Bobrow ,  Karl Edwin Adler ,  Kevin Aaron Roth

IPC分类号： C12Q128

CPC分类号： C12Q1/28 ,  C07C235/20

摘要： A method for enhancing the conversion of a phenol substrate to a product by a peroxidase enzyme comprises the steps of reacting a conjugate comprising a detectably labeled phenol with a peroxidase enzyme in the presence of an enhancing reagent, the enhancing reagent comprising an inorganic salt, an organic compound having the formula wherein when X is B(OH)2, Y is I, or wherein when X is OH, Y is a halogen, or Q-R wherein Q is a linear or branched 1-12 heteroatom alkyl wherein tie heteroatoms are selected from C, N, O, and S, wherein the bonds connecting the heteroatom alkyl chain are single or double, wherein any carbon atom in the heteroatom alkyl chain optionally includes a substituent selected from —OH, —COOH, —NH2, and —SH, and wherein R is selected from —OH, —COOH, —NH2, and —CH3; or mixtures of the inorganic salt and the organic compound. A compound having the formula: wherein R1 is —CH2OH, —COOH, and n is 1-8.

摘要翻译： 通过过氧化物酶增强酚底物转化为产物的方法包括以下步骤：在增强试剂存在下使包含可检测标记的苯酚的缀合物与过氧化物酶反应，所述增强剂包含无机盐， 当X是B（OH）2，Y是I时，或其中当X是OH，Y是卤素或Q 2时，其中Q是直链或支链1-12杂原子烷基，其中连接杂原子选自 C，N，O和S，其中连接杂原子烷基链的键是单或双键，其中杂原子烷基链中的任何碳原子任选地包括选自-OH，-COOH，-NH 2和-SH的取代基， 并且其中R选自-OH，-COOH，-NH 2和-CH 3; 或无机盐和有机化合物的混合物。具有下式的化合物：其中R 1是-CH 2 OH，-COOH，n是1-8。