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公开(公告)号：US6159693A
公开(公告)日：2000-12-12
申请号：US252436
申请日：1999-02-18
CPC分类号： C12Q1/6823 , C12Q1/04 , C12Q1/66 , C12Q1/68 , C12Q1/6827 , Y10S435/81 , C12Q2535/131 , C12Q2521/319 , C12Q2535/107
摘要： This invention discloses methods for detecting specific nucleic acid sequences, interrogating the identity of a specific base within a sequence, and assaying endonuclease and exonuclease activity. DNA or RNA probes are hybridized to target nucleic acid sequences. Probes that are complementary to the target sequence at each base are depolymerized, while probes which differ from the target at the interrogation position are not depolymerized. The nucleic acid detection systems utilize the pyrophosphorolysis reaction catalyzed by various polymerases to produce deoxyribonucleoside triphosphates or ribonucleoside triphosphates. dNTPs are transformed to ATP by the action of NDPK. The ATP produced by these reactions is detected by luciferase or NADH based detection systems.
摘要翻译： 本发明公开了用于检测特定核酸序列，询问序列内特定碱基的同一性以及测定核酸内切酶和核酸外切酶活性的方法。 DNA或RNA探针与靶核酸序列杂交。 与每个碱基上的靶序列互补的探针被解聚，而在询问位置不同于靶的探针没有解聚。 核酸检测系统利用由各种聚合酶催化的焦磷酸解反应产生脱氧核糖核苷三磷酸或核糖核苷三磷酸。 dNTPs通过NDPK的作用转化为ATP。 通过荧光素酶或NADH检测系统检测由这些反应产生的ATP。
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公开(公告)号：US06379898B2
公开(公告)日：2002-04-30
申请号：US09757132
申请日：2001-01-09
IPC分类号： C12Q168
CPC分类号： C12Q1/6823 , C12Q1/04 , C12Q1/66 , C12Q1/68 , C12Q1/6827 , Y10S435/81 , C12Q2535/131 , C12Q2521/319 , C12Q2535/107
摘要： This invention discloses methods, compositions and kits for the detection of extremely low levels of nucleic acid, cells and cellular material in biological samples. The nucleic acid detection systems utilize either the pyrophosphorolysis reaction catalyzed by various polymerases or nuclease digestion coupled with pyrophosphorylation catalyzed by phosphoribosylpyrophosphate synthetase to produce either deoxyribonucleoside triphosphates or ribonucleoside triphosphates. dNTPs are transformed to ATP by the action of nucleoside diphosphate kinase. The ATP produced by these reactions may be detected by luciferase or NADH based detection systems. If more sensitive detection is required, schemes for the amplification of NTPs and dNTPS are provided. A detection system for cells or cellular material in a sample is provided wherein AMP and a high energy phosphate donor added to a sample are converted to ATP by the action of endogenous enzymes, followed by detection of the ATP.
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公开(公告)号：US06335162B1
公开(公告)日：2002-01-01
申请号：US09042287
申请日：1998-03-13
IPC分类号： C12Q168
CPC分类号： C12Q1/6823 , C12Q1/04 , C12Q1/66 , C12Q1/68 , C12Q1/6827 , Y10S435/81 , C12Q2535/131 , C12Q2521/319 , C12Q2535/107
摘要： This invention discloses methods, compositions and kits for the detection of extremely low levels of nucleic acid, cells and cellular material in biological samples. The nucleic acid detection systems utilize either the pyrophosphorolysis reaction catalyzed by various polymerases or nuclease digestion coupled with pyrophosphorylation catalyzed by phosphoribosylpyrophosphate synthetase to produce either deoxyribonucleoside triphosphates or ribonucleoside triphosphates. dNTPs are transformed to ATP by the action of nucleoside diphosphate kinase. The ATP produced by these reactions may be detected by luciferase or NADH based detection systems. If more sensitive detection is required, schemes for the amplification of NTPs and dNTPS are provided. A detection system for cells or cellular material in a sample is provided wherein AMP and a high energy phosphate donor added to a sample are converted to ATP by the action of endogenous enzymes, followed by detection of the ATP.
摘要翻译： 本发明公开了用于检测生物样品中极低水平的核酸，细胞和细胞物质的方法，组合物和试剂盒。 核酸检测系统利用通过各种聚合酶催化的焦磷酸裂解反应或核酸酶消化与磷酸核糖焦磷酸合成酶催化的焦磷酸化反应产生脱氧核糖核苷三磷酸或核糖核苷三磷酸。 dNTPs通过核苷二磷酸激酶的作用转化为ATP。 由这些反应产生的ATP可以通过荧光素酶或基于NADH的检测系统来检测。 如果需要更灵敏的检测，则提供扩增NTPs和dNTPS的方案。 提供样品中的细胞或细胞材料的检测系统，其中通过内源酶的作用将AMP和加入样品的高能量磷酸盐供体转化为ATP，随后检测ATP。
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