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1. 发明申请

US20060088865A1 Methods and compositions related thereto for detecting and identifying distinct species of nucleic acids from causative agents 有权
公开(公告)号：US20060088865A1
公开(公告)日：2006-04-27
申请号：US11246976
申请日：2005-10-07
申请人： Martin Adelson , Melanie Feola , Jason Trama , Eli Mordechai
发明人： Martin Adelson , Melanie Feola , Jason Trama , Eli Mordechai
IPC分类号： C12Q1/70 , C12Q1/68
CPC分类号： C12Q1/705
摘要： Methods are described herein for detecting and identifying distinct species of nucleic acids, in a single container, for example, from a certain genus of infectious agents or otherwise causative agents comprising, for example, providing a forward PCR primer common to a homologous gene region between the distinct species, and providing a reverse PCR primer common to a homologous gene region between the distinct species, to thereby define a PCR target region amongst the species, and providing a first oligonucleotide probe specific to a nucleic acid sequence within the target region that is characteristic of a first species, providing a second oligonucleotide probe specific to a nucleic acid sequence within the target region that is characteristic of a second species, wherein the first and second oligonucleotide probes are each detectably labeled with distinctly different detectable labels, conducting a PCR reaction in the container by means of the primers to amplify the target region amongst the species, and detecting the distinct labels, thereby identifying distinct species of nucleic acids corresponding to distinct species of infectious agents. Methods are preferred, for example, wherein the infectious agent is a member of the Herpesviridae family.

2. 发明授权

US08206930B2 Compositions and methods for detecting Borrelia afzelii 有权
公开(公告)号：US08206930B2
公开(公告)日：2012-06-26
申请号：US13199093
申请日：2011-08-18
申请人： Melanie Feola , Martin Adelson , Eli Mordechai , Lisa Novak
发明人： Melanie Feola , Martin Adelson , Eli Mordechai , Lisa Novak
IPC分类号： C12Q1/68 , C12P19/34 , C07H21/02 , C07H21/04
CPC分类号： C12Q1/689 , Y02A50/57
摘要： Disclosed are oligonucleotides useful in methods for determining whether a sample contains Borrelia afzelii, a spirochete which is a causative agent of Lyme disease in humans. These oligonucleotides, which have nucleotide sequences derived from a coding segment of the gene encoding the p24 gene for the outer surface protein of Borrelia afzelii, are useful as forward and reverse primers for a polymerase chain reaction using nucleic acids from a biological sample as templates, and as probes for detecting any resultant amplicon. Detection of an amplicon indicates the sample contains Borrelia afzelii. Real-time PCR and detection using florescence resonance energy transfer is disclosed.

3. 发明授权

US08026056B2 Compositions and methods for detecting Borrelia afzelii 有权
标题翻译：组织和检测方法
公开(公告)号：US08026056B2
公开(公告)日：2011-09-27
申请号：US12156029
申请日：2008-05-29
申请人： Melanie Feola , Martin Adelson , Eli Mordechai , Lisa Novak
发明人： Melanie Feola , Martin Adelson , Eli Mordechai , Lisa Novak
IPC分类号： C12P19/34 , C12Q1/68 , C07H21/02 , C07H21/04
CPC分类号： C12Q1/689 , Y02A50/57
摘要： Disclosed are oligonucleotides useful in methods for determining whether a sample contains Borrelia afzelii, a spirochete which is a causative agent of Lyme disease in humans. These oligonucleotides, which have nucleotide sequences derived from a coding segment of the gene encoding the p24 gene for the outer surface protein of Borrelia afzelii, are useful as forward and reverse primers for a polymerase chain reaction using nucleic acids from a biological sample as templates, and as probes for detecting any resultant amplicon. Detection of an amplicon indicates the sample contains Borrelia afzelii. Real-time PCR and detection using florescence resonance energy transfer is disclosed.
摘要翻译：公开了用于确定样品是否含有人类的莱姆病的致病因子的螺旋体（Borrelia afzelii）的方法中的寡核苷酸。这些寡核苷酸具有衍生自编码b24基因的基因的编码区段的核苷酸序列，用于使用来自生物样品的核酸作为模板的聚合酶链反应的正向和反向引物，以及用于检测任何所得扩增子的探针。扩增子的检测表明样品含有Borrelia afzelii。公开了使用荧光共振能量转移的实时PCR和检测。

4. 发明申请

US20090275023A1 Compositions and methods for detecting borrelia afzelii 有权
标题翻译：组合和检测方法
公开(公告)号：US20090275023A1
公开(公告)日：2009-11-05
申请号：US12156029
申请日：2008-05-29
申请人： Melanie Feola , Martin Adelson , Eli Mordechai , Lisa Novak
发明人： Melanie Feola , Martin Adelson , Eli Mordechai , Lisa Novak
IPC分类号： C12Q1/68 , C07H21/04
CPC分类号： C12Q1/689 , Y02A50/57
摘要： Disclosed are oligonucleotides useful in methods for determining whether a sample contains Borrelia afzelii, a spirochete which is a causative agent of Lyme disease in humans. These oligonucleotides, which have nucleotide sequences derived from a coding segment of the gene encoding the p24 gene for the outer surface protein of Borrelia afzelii, are useful as forward and reverse primers for a polymerase chain reaction using nucleic acids from a biological sample as templates, and as probes for detecting any resultant amplicon. Detection of an amplicon indicates the sample contains Borrelia afzelii. Real-time PCR and detection using florescence resonance energy transfer is disclosed.
摘要翻译：公开了用于确定样品是否含有人类的莱姆病的致病因子的螺旋体（Borrelia afzelii）的方法中的寡核苷酸。这些寡核苷酸具有衍生自编码b24基因的基因的编码区段的核苷酸序列，用于使用来自生物样品的核酸作为模板的聚合酶链反应的正向和反向引物，以及用于检测任何所得扩增子的探针。扩增子的检测表明样品含有Borrelia afzelii。公开了使用荧光共振能量转移的实时PCR和检测。

5. 发明授权

US08377656B2 Compositions and methods for detecting Cryptococcus neoformans 有权
公开(公告)号：US08377656B2
公开(公告)日：2013-02-19
申请号：US13135022
申请日：2011-06-23
申请人： Melanie Feola , John Entwistle , Martin Adelson , Eli Mordechai
发明人： Melanie Feola , John Entwistle , Martin Adelson , Eli Mordechai
IPC分类号： C12P19/34 , C12Q1/68 , C07H21/04
CPC分类号： C12Q1/6895 , C07H21/04 , C12Q2600/158
摘要： Disclosed are oligonucleotides useful in methods for determining whether a sample contains Cryptococcus neoformans, a causative agent for human cryptococcosis. These oligonucleotides, which have nucleotide sequences derived from a coding segment of the gene encoding the fungal specific transcription factor gene in Cryptococcus neoformans, are useful as forward and reverse primers for a polymerase chain reaction using nucleic acids from a biological sample as templates, and as probes for detecting any resultant amplicon. Detection of an amplicon indicates the sample contains Cryptococcus neoformans. Real-time PCR and detection using florescence resonance energy transfer is disclosed.

6. 发明申请

US20120122092A1 Compositions and methods for detecting Borrelia afzelii 有权
标题翻译：组织和检测方法
公开(公告)号：US20120122092A1
公开(公告)日：2012-05-17
申请号：US13199093
申请日：2011-08-18
申请人： Melanie Feola , Martin Adelson , Eli Mordechai , Lisa Novak
发明人： Melanie Feola , Martin Adelson , Eli Mordechai , Lisa Novak
IPC分类号： C12Q1/68
CPC分类号： C12Q1/689 , Y02A50/57
摘要： Disclosed are oligonucleotides useful in methods for determining whether a sample contains Borrelia afzelii, a spirochete which is a causative agent of Lyme disease in humans. These oligonucleotides, which have nucleotide sequences derived from a coding segment of the gene encoding the p24 gene for the outer surface protein of Borrelia afzelii, are useful as forward and reverse primers for a polymerase chain reaction using nucleic acids from a biological sample as templates, and as probes for detecting any resultant amplicon. Detection of an amplicon indicates the sample contains Borrelia afzelii. Real-time PCR and detection using florescence resonance energy transfer is disclosed.
摘要翻译：公开了用于确定样品是否含有人类的莱姆病的致病因子的螺旋体（Borrelia afzelii）的方法中使用的寡核苷酸。这些寡核苷酸具有衍生自编码b24基因的基因的编码区段的核苷酸序列，用于使用来自生物样品的核酸作为模板的聚合酶链反应的正向和反向引物，以及用于检测任何所得扩增子的探针。扩增子的检测表明样品含有Borrelia afzelii。公开了使用荧光共振能量转移的实时PCR和检测。

7. 发明授权

US07989186B2 Compositions and methods for detecting cryptococcus neoformans 有权
标题翻译：用于检测隐球菌新生隐球菌的组合物和方法
公开(公告)号：US07989186B2
公开(公告)日：2011-08-02
申请号：US12214317
申请日：2008-06-18
申请人： Melanie Feola , Martin Adelson , Eli Mordechai , John Entwistle
发明人： Melanie Feola , Martin Adelson , Eli Mordechai , John Entwistle
IPC分类号： C12P19/34
CPC分类号： C12Q1/6895 , C07H21/04 , C12Q2600/158
摘要： Disclosed are oligonucleotides useful in methods for determining whether a sample contains Cryptococcus neoformans, a causative agent for human cryptococcosis. These oligonucleotides, which have nucleotide sequences derived from a coding segment of the gene encoding the fungal specific transcription factor gene in Cryptococcus neoformans, are useful as forward and reverse primers for a polymerase chain reaction using nucleic acids from a biological sample as templates, and as probes for detecting any resultant amplicon. Detection of an amplicon indicates the sample contains Cryptococcus neoformans. Real-time PCR and detection using florescence resonance energy transfer is disclosed.
摘要翻译：公开了用于确定样品是否含有新隐球菌隐球菌（人类隐球菌病的致病因子）的方法中的寡核苷酸。这些寡核苷酸具有源自编码新型隐球菌中的真菌特异性转录因子基因的基因的编码区的核苷酸序列，可用作使用来自生物样品的核酸作为模板的聚合酶链反应的正向和反向引物，以及用于检测任何所得扩增子的探针。扩增子的检测表明样品含有新型隐球酵母。公开了使用荧光共振能量转移的实时PCR和检测。

8. 发明申请

US20120015362A1 Compositions and methods for detecting cryptococcus neoformans 有权
标题翻译：用于检测隐球菌新生隐球菌的组合物和方法
公开(公告)号：US20120015362A1
公开(公告)日：2012-01-19
申请号：US13135022
申请日：2011-06-23
申请人： Melanie Feola , Martin Adelson , Eli Mordechai , John Entwistle
发明人： Melanie Feola , Martin Adelson , Eli Mordechai , John Entwistle
IPC分类号： C12Q1/68 , C07H21/04
CPC分类号： C12Q1/6895 , C07H21/04 , C12Q2600/158
摘要： Disclosed are oligonucleotides useful in methods for determining whether a sample contains Cryptococcus neoformans, a causative agent for human cryptococcosis. These oligonucleotides, which have nucleotide sequences derived from a coding segment of the gene encoding the fungal specific transcription factor gene in Cryptococcus neoformans, are useful as forward and reverse primers for a polymerase chain reaction using nucleic acids from a biological sample as templates, and as probes for detecting any resultant amplicon. Detection of an amplicon indicates the sample contains Cryptococcus neoformans. Real-time PCR and detection using florescence resonance energy transfer is disclosed.
摘要翻译：公开了用于确定样品是否含有新隐球菌隐球菌（人类隐球菌病的致病因子）的方法中的寡核苷酸。这些寡核苷酸具有源自编码新型隐球菌中的真菌特异性转录因子基因的基因的编码区的核苷酸序列，可用作使用来自生物样品的核酸作为模板的聚合酶链反应的正向和反向引物，以及用于检测任何所得扩增子的探针。扩增子的检测表明样品含有新型隐球酵母。公开了使用荧光共振能量转移的实时PCR和检测。

9. 发明申请

US20090246767A1 Compositions and methods for detecting Cryptococcus neoformans 有权
标题翻译：用于检测新型隐球酵母的组合物和方法
公开(公告)号：US20090246767A1
公开(公告)日：2009-10-01
申请号：US12214317
申请日：2008-06-18
申请人： Melanie Feola , Martin Adelson , Eli Mordechai , John Entwistle
发明人： Melanie Feola , Martin Adelson , Eli Mordechai , John Entwistle
IPC分类号： C12Q1/68 , C07H21/04
CPC分类号： C12Q1/6895 , C07H21/04 , C12Q2600/158
摘要： Disclosed are oligonucleotides useful in methods for determining whether a sample contains Cryptococcus neoformans, a causative agent for human cryptococcosis. These oligonucleotides, which have nucleotide sequences derived from a coding segment of the gene encoding the fungal specific transcription factor gene in Cryptococcus neoformans, are useful as forward and reverse primers for a polymerase chain reaction using nucleic acids from a biological sample as templates, and as probes for detecting any resultant amplicon. Detection of an amplicon indicates the sample contains Cryptococcus neoformans. Real-time PCR and detection using florescence resonance energy transfer is disclosed.
摘要翻译：公开了用于确定样品是否含有新隐球菌隐球菌（人类隐球菌病的致病因子）的方法中的寡核苷酸。这些寡核苷酸具有源自编码新型隐球菌中的真菌特异性转录因子基因的基因的编码区的核苷酸序列，可用作使用来自生物样品的核酸作为模板的聚合酶链反应的正向和反向引物，以及用于检测任何所得扩增子的探针。扩增子的检测表明样品含有新型隐球酵母。公开了使用荧光共振能量转移的实时PCR和检测。

10. 发明申请

US20060127934A1 Oligonucleotides useful in methods for detecting and characterizing Aspergillus fumigatus 审中-公开
公开(公告)号：US20060127934A1
公开(公告)日：2006-06-15
申请号：US11299362
申请日：2005-12-09
申请人： Jason Trama , Martin Adelson , Eli Mordechai
发明人： Jason Trama , Martin Adelson , Eli Mordechai
IPC分类号： C12Q1/68 , C07H21/04
CPC分类号： C12Q1/6895 , C12Q1/689 , C12Q2561/113 , C12Q2565/301 , C12Q2600/156
摘要： Oligonucleotides and methods for using these oligonucleotides in the detection of Aspergillus fumigatus are disclosed. Aspergillus fumigatus is the causative agent for medical conditions including invasive aspergillosis. The oligonucleotides of the invention have nucleotide sequences derived from the gene encoding the cytochrome P450 14 alpha-sterol demethylase (i.e., the Cyp51A protein) of Aspergillus fumigatus. The oligonucleotides of the invention include forward primers and reverse primers which in combination are capable of priming the synthesis of amplicons specific to cyp51A in polymerase chain reactions using nucleic acids isolated from Aspergillus fumigatus as templates. The oligonucleotides of the invention also include probes capable of detecting these cyp51A-specific amplicons. Thus, a biological sample is tested for the presence of Aspergillus fumigatus by isolating nucleic acid from the sample, conducting a polymerase chain reaction in a mixture containing this nucleic acid and these forward and reverse primers, and then determining, using an oligonucleotide probe, whether an amplicon is produced in the mixture, wherein detection of the amplicon indicates the presence of Aspergillus fumigatus in the sample. The oligonucleotides of the invention also include primers for nucleotide sequencing reactions to determine whether an isolate of Aspergillus fumigatus is more tolerant than wild-type Aspergillus fumigatus to a triazole, which is a compound commonly used as an antifungal drug. Specifically, a strand of a cyp51A-specific amplicon is at least partially sequenced using a nucleotide sequencing primer in a primer extension reaction. Identification of mutations giving rise to amino acid substitutions at positions 54, 138, 220, and 448 of the amino acid sequence of the wild-type Cyp51A protein indicates that the isolate of Aspergillus fumigatus from which the amplicon is derived exhibits decreased susceptibility to at least one triazole.

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