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1.发明授权
公开(公告)号：US5561044A
公开(公告)日：1996-10-01
申请号：US398305
申请日：1995-03-03
申请人： George T. Walker , James G. Nadeau , Patricia A. Spears , Colleen M. Nycz , Daryl D. Shank , James L. Schram , Stewart R. Jurgensen
发明人： George T. Walker , James G. Nadeau , Patricia A. Spears , Colleen M. Nycz , Daryl D. Shank , James L. Schram , Stewart R. Jurgensen
CPC分类号： C12Q1/689 , C12Q1/6851 , C12Q2600/16
摘要： Primers and methods for adapter-mediated multiplex amplification of the IS6110 insertion element of Mycobacterium tuberculosis (M.tb) and the 16S ribosomal gene of Mycobacterium tuberculosis, useful for simultaneously detecting and/or identifying species of the M. tuberculosis complex and other clinically relevant Mycobacterium species. Multiplex Strand Displacement Amplification (SDA) is used in a single amplification reaction which is capable of simultaneously identifying M. tuberculosis and providing a screen for substantially all of the clinically relevant species of Mycobacteria. Also disclosed are methods for adapter-mediated multiplex amplification of multiple target sequences and a single internal control sequence for determination of sample efficacy or quantitation of the targets. In a preferred embodiment, an internal control sequence is included in the amplification reaction and coamplified with the IS6110 and 16S target sequences as an indication of sample amplification activity or to quantitate the initial amount of target sequences in the sample.
摘要翻译： 针对结核分枝杆菌（M.tb）的IS6110插入元件和结核分枝杆菌的16S核糖体基因的适配器介导的多重扩增的引物和方法，可用于同时检测和/或鉴定结核分枝杆菌复合物和其他临床相关的物种 分枝杆菌种。 多重链位移扩增（SDA）用于能够同时鉴定结核分枝杆菌并为基本上所有临床相关分枝杆菌物种提供筛选的单一扩增反应。 还公开了用于多重靶序列的衔接子介导的多重扩增的方法和用于确定靶的样品功效或定量的单个内部控制序列的方法。 在优选的实施方案中，内部对照序列包括在扩增反应中并与IS6110和16S靶序列共扩增，作为样品扩增活性的指示或定量样品中靶序列的初始量。
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公开(公告)号：US5518884A
公开(公告)日：1996-05-21
申请号：US203534
申请日：1994-02-28
申请人： Patricia A. Spears , Daryl D. Shank
发明人： Patricia A. Spears , Daryl D. Shank
CPC分类号： C12Q1/689
摘要： Oligonucleotide probes and primers which exhibit M. kansasii-specificity in nucleic acid hybridization assays and in nucleic acid amplification reactions are provided. The full-length M. kansasii-specific sequence, identified herein as clone MK7, is 493 base pairs in length and has a GC content of 63%. Several M. kansasii-specific subsequences of MK7 are also provided. The probes and primers are useful in assays for species-specific detection and identification of M. kansasii.
摘要翻译： 提供了在核酸杂交测定和核酸扩增反应中表现出堪萨斯样菌株特异性的寡核苷酸探针和引物。 本文鉴定为克隆MK7的全长M.堪萨斯特异性序列长度为493个碱基对，GC含量为63％。 还提供了MK7的几种堪萨斯特异性亚序列。 探针和引物可用于物种特异性检测和鉴定堪萨斯统计分析。
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公开(公告)号：US5500341A
公开(公告)日：1996-03-19
申请号：US308892
申请日：1994-09-19
申请人： Patricia A. Spears
发明人： Patricia A. Spears
CPC分类号： C12Q1/689
摘要： M. kansasii-specific oligonucleotide amplification primers and methods for detecting and identifying M. kansasii nucleic acids using the amplification primers. One hundred percent of the clinical and environmental M. kanasii isolates tested were positive in amplification assays using the inventive amplification primers, with no cross-reactivity in other species of mycobacteria or closely related non-mycobacteria.
摘要翻译： 堪萨斯体特异性寡核苷酸扩增引物以及使用扩增引物检测和鉴定堪萨斯假单胞菌核酸的方法。 使用本发明的扩增引物，在扩增测定中测试的100％的临床和环境卡氏菌分离物是阳性的，在其他分枝杆菌或密切相关的非分枝杆菌中没有交叉反应性。
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公开(公告)号：US5494796A
公开(公告)日：1996-02-27
申请号：US275225
申请日：1994-07-14
CPC分类号： C12Q1/689
摘要： Genus- and species-specific oligonucleotide probes derived from the M. paratuberculosis 70 kD heat shock protein gene sequence. The probes are useful for detecting Mycobacteria and for identifying specific species of Mycobacteria.
摘要翻译： 来源于副结核分枝杆菌70 kD热休克蛋白基因序列的属和物种特异性寡核苷酸探针。 探针可用于检测分枝杆菌和鉴定分枝杆菌的特定物种。
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公开(公告)号：US5470723A
公开(公告)日：1995-11-28
申请号：US111076
申请日：1993-08-24
申请人： George T. Walker , James G. Nadeau , Patricia A. Spears , Colleen M. Nycz , Daryl D. Shank , James L. Schram , Stewart R. Jurgensen
发明人： George T. Walker , James G. Nadeau , Patricia A. Spears , Colleen M. Nycz , Daryl D. Shank , James L. Schram , Stewart R. Jurgensen
CPC分类号： C12Q1/689 , C12Q1/6851 , C12Q2600/16
摘要： Primers and methods for adapter-mediated multiplex amplification of the IS6110 insertion element of Mycobacterium tuberculosis (M.tb) and the 16S ribosomal gene of Mycobacterium tuberculosis, useful for simultaneously detecting and/or identifying species of the M. tuberculosis complex and other clinically relevant Mycobacterium species. Multiplex Strand Displacement Amplification (SDA) is used in a single amplification reaction which is capable of simultaneously identifying M. tuberculosis and providing a screen for substantially all of the clinically relevant species of Mycobacteria. Also disclosed are methods for adapter-mediated multiplex amplification of multiple target sequences and a single internal control sequence for determination of sample efficacy or quantitation of the targets. In a preferred embodiment, an internal control sequence is included in the amplification reaction and coamplified with the IS6110 ant 16S target sequences as an indication of sample amplification activity or to quantitate the initial amount of target sequences in the sample.
摘要翻译： 针对结核分枝杆菌（M.tb）的IS6110插入元件和结核分枝杆菌的16S核糖体基因的适配器介导的多重扩增的引物和方法，可用于同时检测和/或鉴定结核分枝杆菌复合物和其他临床相关的物种 分枝杆菌种。 多重链位移扩增（SDA）用于能够同时鉴定结核分枝杆菌并为基本上所有临床相关分枝杆菌物种提供筛选的单一扩增反应。 还公开了用于多重靶序列的衔接子介导的多重扩增的方法和用于确定靶的样品功效或定量的单个内部控制序列的方法。 在优选的实施方案中，扩增反应中包含内部控制序列并与IS6110蚂蚁16S靶序列共扩增，作为样品扩增活性的指示或定量样品中靶序列的初始量。
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公开(公告)号：US5814490A
公开(公告)日：1998-09-29
申请号：US661507
申请日：1996-06-11
申请人： Patricia A. Spears
发明人： Patricia A. Spears
CPC分类号： C12Q1/689
摘要： Amplification primers useful in assays for species-specific detection of a target sequence in the cryptic plasmid of C. trachomatis are described. The primers of the invention amplify a target in the region of nucleotides 2219-2366 of the cryptic plasmid sequence, and the target binding sequences disclosed may be adapted for use in amplification primers for a variety of amplification reactions.
摘要翻译： 描述了可用于沙眼衣原体隐蔽质粒中靶序列物种特异性检测试验的扩增引物。 本发明的引物扩增了隐匿性质粒序列的核苷酸2219-2366区域中的靶，并且所公开的靶结合序列可适用于多种扩增反应的扩增引物。
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7.发明授权Selective detection of mycobacteria by nucleicacid probes derived from Mycobacterium kansasii 失效通过源自堪萨斯分枝杆菌+ B的核酸探针选择性检测分枝杆菌
公开(公告)号：US5352580A
公开(公告)日：1994-10-04
申请号：US889651
申请日：1993-05-26
申请人： Patricia A. Spears , Daryl D. Shank
发明人： Patricia A. Spears , Daryl D. Shank
CPC分类号： C12Q1/689
摘要： Oligonucleotide probes derived from Mycobacteria Kansasii capable of selectively hybridizing to Mycobacteria nucleic acid are disclosed. The oligonucleotide probe is selected from the group consisting of: (a) an oligonucleotide probe consisting essentially of the DNA sequence given herein as SEQ ID NO:1 (MK14); (b) oligonucleotide probes comprising fragments of MK14 which retain the capability of MK14 of selectively hybridizing to Mycobacteria nucleic acid; (c) oligonucleotide probes which hybridize to MK14 and are capable of selectively hybridizing to Mycobacteria nucleic acid; and (d) oligonucleotide probes which are complementary to any of the foregoing and are capable of selectively hybridizing to Mycobacteria nucleic acid. The probes are useful in nucleic acid amplification and hybridization assays for genus-specific detection of the mycobacteria, specific detection of M. kansasii and specific detection of the slow-growing mycobacteria. Methods of using the probes to detect and amplify Mycobacteria nucleic acid and kits containing the same are also disclosed.
摘要翻译： 公开了衍生自能够与分枝杆菌核酸选择性杂交的堪萨斯分枝杆菌的寡核苷酸探针。 寡核苷酸探针选自：（a）基本上由本文给出的DNA序列（SEQ ID NO：1（MK14））组成的寡核苷酸探针; （b）包含保留与分枝杆菌核酸选择性杂交的MK14能力的MK14片段的寡核苷酸探针; （c）与MK14杂交并且能够与分枝杆菌核酸选择性杂交的寡核苷酸探针; 和（d）与前述任何一个互补并且能够与分枝杆菌核酸选择性杂交的寡核苷酸探针。 该探针可用于核酸扩增和杂交测定，用于分类细菌的属特异性检测，堪萨斯假单胞菌的特异性检测和缓慢生长的分枝杆菌的特异性检测。 还公开了使用探针检测和扩增分枝杆菌核酸的方法以及含有其的试剂盒。
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