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1. 发明申请

WO1995003426A2 METHOD FOR NUCLEIC ACID AMPLIFICATION BY TRANSCRIPTION USING DISPLACEMENT, AND REAGENTS AND KIT THEREFOR 审中-公开
标题翻译：通过使用位移转录进行核酸扩增的方法及其试剂和试剂盒
公开(公告)号：WO1995003426A2
公开(公告)日：1995-02-02
申请号：PCT/FR1994000935
申请日：1994-07-26
申请人： BIO MERIEUX , CLEUZIAT, Philippe , GUILLOU-BONNICI, Françoise , MALLET, François , LEVASSEUR, Pierre
发明人： BIO MERIEUX
IPC分类号： C12Q01/68
CPC分类号： C12Q1/6853 , C12Q1/6844 , C12Q1/6846 , C12Q2531/119 , C12Q2521/107 , C12Q2521/101 , C12Q2537/1373
摘要： A method for amplifying a target nucleic acid sequence by providing a polynucleotide such as VII including the sequence to be amplified as well as an RNA polymerase promoter sequence, and contacting said polynucleotide, in the presence of a system having RNA polymerase activity, RNA-dependent DNA polymerase activity, and DNA-dependent DNA polymerase activity, and capable of strand displacement, with a set of primers includinf a primer such as A capable of hybridising with a segment complementary to a portion of the sequence to be amplified, wherein A includes an upstream RNA polymerase promoter sequence then an arbitrary sequence, a primer such as D contains said RNA polymerase promoter sequence, and a primer such as C contains said arbitrary sequence. The elongation product of D is displaced by the elongation product of C, and the elongation product of D is thus obtained in the form of single-stranded VIII. Hybridisation of A on VIII followed by elongation forms double-stranded product X which can undergo transcription, with amplification, to give single-stranded RNA VII bis. An analogous system of 3 primers B, E and F similary produces single-stranded RNA VII from VII bis. Hence the amplification method, which can operate isothermally, is cyclic.
摘要翻译：通过提供包括待扩增的序列的多核苷酸，以及RNA聚合酶启动子序列，并且在具有RNA聚合酶活性的系统存在下使所述多核苷酸与RNA聚合酶活性相接触来扩增靶核酸序列的方法 DNA聚合酶活性和DNA依赖性DNA聚合酶活性，并且能够使用一组引物，包括能够与与待扩增的序列的一部分互补的区段杂交的引物如A的引物，其中A包括上游RNA聚合酶启动子序列然后是任意序列，引物如D含有所述RNA聚合酶启动子序列，引物如C包含所述任意序列。 D的伸长率由C的伸长率移位，因此以单链VIII的形式得到D的伸长率。 A对VIII的杂交，然后延伸形成双链产物X，其可经历转录，扩增，得到单链RNA VII bis。 3个引物B，E和F的类似系统同样产生VII bis的单链RNA VII。因此，可以等温操作的扩增方法是循环的。

2. 发明申请

WO1998005766A1 TARGET NUCLEIC ACID SEQUENCE AMPLIFICATION METHOD 审中-公开
标题翻译：目标核酸序列放大方法
公开(公告)号：WO1998005766A1
公开(公告)日：1998-02-12
申请号：PCT/FR1997001445
申请日：1997-08-01
申请人： BIO MERIEUX , GUILLOU-BONNICI, Françoise , DEFRANC, Eric , HOANG, Antoine , LAAYOUN, Ali , LHOMME, Jean , TREVISIOL, Emmanuelle
发明人： BIO MERIEUX
IPC分类号： C12N15/10
CPC分类号： C12P19/34 , C07H19/06 , C07H21/00 , C12N15/10 , C12P19/30 , C12Q1/686 , C12Q2527/125 , C12Q2525/101
摘要： A target nucleic acid sequence amplification method wherein at least the target nucleic acid sequence, at least one oligonucleotide primer specific for the target sequence, one or more enzymatic activities and nucleotides are provided, and the target sequence is amplified under conditions suitable in particular for said enzymatic or activity or activities, is disclosed. According to the method, at least one of the nucleotides is a prefunctionalised nucleotide that differs from the other nucleotides at least in that it includes at least one unprotected reactive covalence function in at least one predetermined site of the base of said nucleotide, in order to give a prefunctionalised amplification product including at least one such prefunctionalised nucleotide. Nucleotides for carrying out the method are also disclosed.
摘要翻译：一种目标核酸序列扩增方法，其中至少提供了靶核酸序列，至少一种针对靶序列特异性的寡核苷酸引物，一种或多种酶活性和核苷酸，并且靶序列在适合于所述酶或活性或活性。根据该方法，至少一个核苷酸是与其它核苷酸不同的预功能化核苷酸，至少其中包括至少一个在所述核苷酸碱基的至少一个预定位点的未保护的反应性共价功能，以便给出包含至少一种这样的预功能化核苷酸的前功能化扩增产物。还公开了用于实施该方法的核苷酸。

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