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    • 3. 发明申请
      WO9940224A9 DIRECT DETECTION OF RNA MEDIATED BY REVERSE TRANSCRIPTASE LACKING RNASE H FUNCTION 审中-公开
    • DIRECT DETECTION OF RNA MEDIATED BY REVERSE TRANSCRIPTASE LACKING RNASE H FUNCTION
    • 直接检测由逆转录酶介导的RNA缺失RNASE H功能
    • WO9940224A9
    • 1999-10-21
    • PCT/US9902382
    • 1999-02-03
    • DIGENE CORP
    • DE LA ROSA ABEL , COLLIER CLAYTON D
    • C12Q1/68
    • C12Q1/6804 , C12Q2521/107
    • Disclosed is a method of detecting RNA molecules of interest in which reverse transcription primers unique to the RNA molecule of interest are used for reverse transcribing the RNA with a reverse transcriptase lacking RNAse H function and the resulting RNA/DNA hybrid is detected with an antibody specific for RNA/DNA hybrids. This method can be used to detect the presence of one or many specific RNA molecules which may be present in a sample, including RNA from different organisms (such as viruses, bacteria, fungi, plants, and animals), or RNA indicative of an infection, a disease state, or predisposition to a disease in an animal. The specificity of detection is increased relative to current detection methods involving probe hybridization since the reverse transcription primers are shorter and less subject to non-specific hybridization. Specificity of the disclosed method can also be increased by using a thermostable reverse transcriptase and performing reverse transcription at a high temperature. The disclosed method can also be used to detect reverse transcriptase activity in a sample and to identify inhibitors of reverse transcriptase. Also disclosed is a method for sequencing target RNA molecules using reverse transcriptase lacking an RNAse H function.
    • 本发明公开了一种检测感兴趣的RNA分子的方法，其中将RNA感兴趣的RNA分子特有的逆转录引物用于逆转录RNA，所述反转录酶缺乏RNA酶H功能，并且所得到的RNA / DNA杂合物用抗体特异性 用于RNA / DNA杂交体。 该方法可用于检测可能存在于样品中的一种或多种特定RNA分子的存在，包括来自不同生物（例如病毒，细菌，真菌，植物和动物）的RNA或指示感染的RNA ，疾病状态或动物疾病的倾向。 相对于涉及探针杂交的当前检测方法，检测的特异性增加，因为逆转录引物较短且较少受非特异性杂交。 所公开的方法的特异性也可以通过使用耐热逆转录酶并在高温下进行逆转录来增加。 所公开的方法也可用于检测样品中的逆转录酶活性并鉴定逆转录酶的抑制剂。 还公开了使用缺乏RNA酶H功能的逆转录酶对靶RNA分子进行测序的方法。
    • 基本信息 添加关注 加入工作空间 PDF全文 PDF下载 全文下载 相似专利 双屏查看 权利要求书 说明书全文 Global Dossier Espacenet 法律信息 同族专利 专利引用
    • 7. 发明专利
      CA2320102A1 DIRECT DETECTION OF RNA MEDIATED BY REVERSE TRANSCRIPTASE LACKING RNASE H FUNCTION 未知
    • DIRECT DETECTION OF RNA MEDIATED BY REVERSE TRANSCRIPTASE LACKING RNASE H FUNCTION
    • CA2320102A1
    • 1999-08-12
    • CA2320102
    • 1999-02-03
    • DIGENE CORP
    • COLLIER CLAYTON D , DE LA ROSA ABEL
    • C12Q1/68 , C12Q1/6804
    • Disclosed is a method of detecting RNA molecules of interest in which reverse transcription primers unique to the RNA molecule of interest are used for reverse transcribing the RNA with a reverse transcriptase lacking RNAse H function and the resulting RNA/DNA hybrid is detected with an antibody specific for RNA/DNA hybrids. This method can be used to detect the presence of one or many specific RNA molecules which may be present in a sample, including RNA from different organisms (such as viruses, bacteria, fungi, plants, and animals), or RNA indicative of an infection, a disease state, or predisposition to a disease in an animal. The specificity of detection is increased relative to current detection methods involving probe hybridization since the reverse transcription primers are shorter and less subject to nonspecific hybridization. Specificity of the disclosed method can also be increased by using a thermostable reverse transcriptase and performing reverse transcription at a high temperature. The disclosed method can also be used to detect reverse transcriptase activity in a sample and to identify inhibitors of reverse transcriptase. Also disclosed is a method for sequencing target RNA molecules using reverse transcriptase lacking an RNAse H function.
    • 基本信息 添加关注 加入工作空间 PDF全文 PDF下载 全文下载 相似专利 双屏查看 权利要求书 说明书全文 Espacenet 法律信息 同族专利 专利引用
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