公开(公告)号：US20110104718A1

公开(公告)日：2011-05-05

申请号：US12917630

申请日：2010-11-02

申请人： Galla Chandra Rao ,  Christopher Larson ,  Madeline Repollet ,  Herman Rutner ,  Leon W.M.M. Terstappen ,  Shawn Mark O'Hara ,  Steven Gross

发明人： Galla Chandra Rao ,  Christopher Larson ,  Madeline Repollet ,  Herman Rutner ,  Leon W.M.M. Terstappen ,  Shawn Mark O'Hara ,  Steven Gross

IPC分类号： G01N33/574 ,  G01N33/566

CPC分类号： G01N33/574 ,  G01N33/54326 ,  Y10S436/813 ,  Y10T436/101666 ,  Y10T436/25125 ,  Y10T436/25375

摘要： The methods and reagents described in this invention are used to analyze circulating tumor cells, clusters, fragments, and debris. Analysis is performed with a number of platforms, including flow cytometry and the CellSpotter® fluorescent microscopy imaging system. Analyzing damaged cells has shown to be important. However, there are two sources of damage: in vivo and in vitro. Damage in vivo occurs by apoptosis, necrosis, or immune response. Damage in vitro occurs during sample acquisition, handling, transport, processing, or analysis. It is therefore desirable to confine, reduce, eliminate, or at least qualify in vitro damage to prevent it from interfering in analysis. Described herein are methods to diagnose, monitor, and screen disease based on circulating rare cells, including malignancy as determined by CTC, clusters, fragments, and debris. Also provided are kits for assaying biological specimens using these methods.

摘要翻译： 本发明描述的方法和试剂用于分析循环肿瘤细胞，簇，碎片和碎片。 使用许多平台进行分析，包括流式细胞术和CellSpotter®荧光显微镜成像系统。 分析损伤的细胞已被证明是重要的。 然而，有两个损害来源：体内和体外。 体内损伤是通过细胞凋亡，坏死或免疫应答发生的。 在样品采集，处理，运输，加工或分析过程中发生体外损伤。 因此，期望限制，减少，消除或至少限定体外损伤，以防止其干扰分析。 本文描述了基于循环稀有细胞（包括由CTC，簇，碎片和碎片确定的恶性肿瘤）来诊断，监测和筛选疾病的方法。 还提供了使用这些方法测定生物样品的试剂盒。

公开(公告)号：US07863012B2

公开(公告)日：2011-01-04

申请号：US10780399

申请日：2004-02-17

申请人： Galla Chandra Rao ,  Christopher Larson ,  Madeline Repollet ,  Herman Rutner ,  Leon W. M. M. Terstappen ,  Shawn Mark O'Hara ,  Steven Gross

发明人： Galla Chandra Rao ,  Christopher Larson ,  Madeline Repollet ,  Herman Rutner ,  Leon W. M. M. Terstappen ,  Shawn Mark O'Hara ,  Steven Gross

IPC分类号： A01N1/02 ,  G01N33/574 ,  G01N33/553 ,  G01N33/00

CPC分类号： G01N33/574 ,  G01N33/54326 ,  Y10S436/813 ,  Y10T436/101666 ,  Y10T436/25125 ,  Y10T436/25375

摘要： The methods and reagents described in this invention are used to analyze circulating tumor cells, clusters, fragments, and debris. Analysis is performed with a number of platforms, including flow cytometry and the CellSpotter® fluorescent microscopy imaging system. Analyzing damaged cells has shown to be important. However, there are two sources of damage: in vivo and in vitro. Damage in vivo occurs by apoptosis, necrosis, or immune response. Damage in vitro occurs during sample acquisition, handling, transport, processing, or analysis. It is therefore desirable to confine, reduce, eliminate, or at least qualify in vitro damage to prevent it from interfering in analysis. Described herein are methods to diagnose, monitor, and screen disease based on circulating rare cells, including malignancy as determined by CTC, clusters, fragments, and debris. Also provided are kits for assaying biological specimens using these methods.

摘要翻译： 本发明描述的方法和试剂用于分析循环肿瘤细胞，簇，碎片和碎片。 使用许多平台进行分析，包括流式细胞术和CellSpotter®荧光显微镜成像系统。 分析损伤的细胞已被证明是重要的。 然而，有两个损害来源：体内和体外。 体内损伤是通过细胞凋亡，坏死或免疫应答发生的。 在样品采集，处理，运输，加工或分析过程中发生体外损伤。 因此，期望限制，减少，消除或至少限定体外损伤，以防止其干扰分析。 本文描述了基于循环稀有细胞（包括由CTC，簇，碎片和碎片确定的恶性肿瘤）来诊断，监测和筛选疾病的方法。 还提供了使用这些方法测定生物样品的试剂盒。

公开(公告)号：US08329422B2

公开(公告)日：2012-12-11

申请号：US12917630

申请日：2010-11-02

申请人： Galla Chandra Rao ,  Christopher Larson ,  Madeline Repollet ,  Herman Rutner ,  Leon W. M. M. Terstappen ,  Shawn Mark O'Hara ,  Steven Gross

发明人： Galla Chandra Rao ,  Christopher Larson ,  Madeline Repollet ,  Herman Rutner ,  Leon W. M. M. Terstappen ,  Shawn Mark O'Hara ,  Steven Gross

IPC分类号： G01N33/574 ,  G01N33/553 ,  G01N1/18

CPC分类号： G01N33/574 ,  G01N33/54326 ,  Y10S436/813 ,  Y10T436/101666 ,  Y10T436/25125 ,  Y10T436/25375

摘要： The methods and reagents described in this invention are used to analyze circulating tumor cells, clusters, fragments, and debris. Analysis is performed with a number of platforms, including flow cytometry and the CELLSPOTTER® fluorescent microscopy imaging system. Analyzing damaged cells has shown to be important. However, there are two sources of damage: in vivo and in vitro. Damage in vivo occurs by apoptosis, necrosis, or immune response. Damage in vitro occurs during sample acquisition, handling, transport, processing, or analysis. It is therefore desirable to confine, reduce, eliminate, or at least qualify in vitro damage to prevent it from interfering in analysis. Described herein are methods to diagnose, monitor, and screen disease based on circulating rare cells, including malignancy as determined by CTC, clusters, fragments, and debris. Also provided are kits for assaying biological specimens using these methods.

摘要翻译： 本发明描述的方法和试剂用于分析循环肿瘤细胞，簇，碎片和碎片。 使用许多平台进行分析，包括流式细胞术和CELLSPOTTER®荧光显微镜成像系统。 分析损伤的细胞已被证明是重要的。 然而，有两个损害来源：体内和体外。 体内损伤是通过细胞凋亡，坏死或免疫应答发生的。 在样品采集，处理，运输，加工或分析过程中发生体外损伤。 因此，期望限制，减少，消除或至少限定体外损伤，以防止其干扰分析。 本文描述了基于循环稀有细胞（包括由CTC，簇，碎片和碎片确定的恶性肿瘤）来诊断，监测和筛选疾病的方法。 还提供了使用这些方法测定生物样品的试剂盒。

公开(公告)号：US20130196318A1

公开(公告)日：2013-08-01

申请号：US13641480

申请日：2011-04-15

申请人： Shawn Mark O'Hara ,  Daniel Zweitzig

发明人： Shawn Mark O'Hara ,  Daniel Zweitzig

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6888 ,  C12Q1/04 ,  C12Q1/25 ,  C12Q1/42 ,  C12Q1/48

摘要： The present invention relates generally to the field of detection of microorganisms, in particular detection of bacteria, to methods for measuring enzyme activity, such as DNA polymerase activity, and particularly relates to such methods performed on microbial crude lysates, useful for determining microbial enzyme activities which can be linked to amplification signal generators such as real-time Polymerase Chain Reaction (PCR) techniques, thereby enabling determination of microbial pathogens in samples such as unpurified blood and other body fluids. This invention also relates to reagents for use in such methods, and to test kits comprising such reagents for carrying out the methods.

摘要翻译： 本发明一般涉及微生物的检测领域，特别是细菌的检测方法，用于测量酶活性的方法，例如DNA聚合酶活性，特别涉及用于测定微生物酶活性的微生物粗裂解物上进行的这些方法 其可以与诸如实时聚合酶链式反应（PCR）技术的放大信号发生器相关联，从而能够测定样品中的微生物病原体，例如未纯化的血液和其它体液。 本发明还涉及用于这些方法的试剂，以及用于测试试剂盒，其包含用于实施该方法的这种试剂。

公开(公告)号：US20120077684A1

公开(公告)日：2012-03-29

申请号：US12809090

申请日：2008-12-26

申请人： Shawn Mark O'Hara

发明人： Shawn Mark O'Hara

IPC分类号： C40B30/00 ,  C12Q1/68

CPC分类号： C12Q1/689

摘要： The present invention provides methods, compositions and diagnostic kits for the detection of Staphylococcus Aureus (SA) and antibiotic resistant forms and variants thereof, such as methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Staphylococcus aureus (VRSA), mupirocin-resistant Staphylococcus aureus (mupSA), and the like, in a sample population. The invention preferably involves the improvements of bacterial sampling by means of SA enrichment, followed by SA cell disruption and amplification procedures incorporating the use of multiplex assays for SA specific genes, such as mecA and coagulase negative Staphylococci (CONS) specific genes such as tufA, for SA identification and identification of its known species. This provides means for controlling for the thirty or more known CONS species in assessing SA samples, especially those CONS species that may carry antibiotic resistance genes, such as SCCmec.

摘要翻译： 本发明提供用于检测金黄色葡萄球菌（SA）和抗生素抗性形式及其变体的方法，组合物和诊断试剂盒，例如耐甲氧西林金黄色葡萄球菌（MRSA），耐万古霉素金黄色葡萄球菌（VRSA），耐多西抗素 样本群体中的金黄色葡萄球菌（mupSA）等。 本发明优选地涉及通过SA富集来改进细菌取样，随后是SA细胞破碎和扩增程序，其包括使用针对SA特异性基因的多重测定法，例如mecA和凝固酶阴性葡萄球菌（CONS）特异性基因，例如tufA， 用于SA识别和鉴定其已知物种。 这提供了在评估SA样品，特别是可携带抗生素抗性基因的那些CONS物种（例如SCCmec）时，控制三十种以上已知的CONS物质的手段。

公开(公告)号：US20100062426A1

公开(公告)日：2010-03-11

申请号：US12092626

申请日：2005-11-03

申请人： Shawn Mark O'Hara ,  Denis Smimov ,  Daniel R. Zweitzig

发明人： Shawn Mark O'Hara ,  Denis Smimov ,  Daniel R. Zweitzig

IPC分类号： C12Q1/68

CPC分类号： G01N33/57415 ,  G01N33/574

摘要： A method for assessing tumor progression is described by assessing AGR2 and/or TFF3 expression in a biological sample after induction of a physiological stress, such as hypoxia or serum deprivation in an enriched sample. Assessing the role of these indicators and their expression levels in an enriched CTC sample provides diagnostic and prognositic information on a patient. This method is also useful as a pharmatool in drug discovery.

摘要翻译： 通过在诱导生理应激（例如富集样品中的缺氧或血清剥夺）之后评估生物样品中的AGR2和/或TFF3表达来描述评估肿瘤进展的方法。 评估这些指标及其在富集的CTC样品中的表达水平的作用提供了对患者的诊断和预后信息。 这种方法也可用作药物发现的药剂。

公开(公告)号：US20140186828A1

公开(公告)日：2014-07-03

申请号：US13977719

申请日：2011-12-27

申请人： Shawn Mark O'Hara

发明人： Shawn Mark O'Hara

IPC分类号： C12Q1/68

CPC分类号： C12Q1/689 ,  C12Q1/6806 ,  C12Q2600/106

摘要： The present invention relates to novel methods, and kits, for selectively excluding dead cells from a mixture containing live and dead cells, such as microbe cells in clinical samples, blood products, medical/biotechnology products and food products where subsequent interrogation of the selected live cells are an indicator of the presence of microbe viability. In particular, the invention relates to improved methods for performing direct nucleic acid amplification techniques such as Polymerase Chain Reaction (PCR) and isothermal techniques in blood and other body fluids, for correlation with microbe cell viability from Bacteremia and Fungemia samples. The improved methods provided by the invention are particularly advantageous for the diagnosis of septicemia and to determine pathological conditions in all other normally sterile body fluids.

摘要翻译： 本发明涉及用于从含有活细胞和死细胞的混合物中选择性排除死细胞的新方法和试剂盒，例如临床样品中的微生物细胞，血液制品，医疗/生物技术产品和食品，随后询问所选择的活 细胞是微生物活力存在的指标。 特别地，本发明涉及用于进行直接核酸扩增技术的改进方法，例如聚合酶链式反应（PCR）和血液和其他体液中的等温技术，用于与菌血症和Fungemia样品的微生物细胞活力相关。 本发明提供的改进方法对于诊断败血症和确定所有其它正常无菌体液中的病理状况是特别有利的。

公开(公告)号：US20110111399A1

公开(公告)日：2011-05-12

申请号：US12809086

申请日：2008-12-23

申请人： Shawn Mark O'Hara ,  Mark James Kopnitsky

发明人： Shawn Mark O'Hara ,  Mark James Kopnitsky

IPC分类号： C12Q1/68

CPC分类号： C12Q1/689 ,  C12Q1/686

摘要： Methods and compositions, including diagnostic kits, for the detection of Staphylococcus Aureus (SA) and clinically important antibiotic resistant forms thereof, such as methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Staphylococcus aureus (VRSA), mupirocin-resistant Staphylococcus aureus (mupSA), and the like, from individuals in a sample population are disclosed Also disclosed are cost effective methods and kits for bacterial sampling and analysis via inherent and expeditious SA cell disruption methods followed by Direct PCR, circumventing the need, expense and contamination πsks associated with DNA isolation methods These improved methods in conjunction with SA prevalence analysis are applied so as to eliminate the approximately 70% of samples in the human population which do not carry SA (SA negative), followed by a second more costly test for antibiotic resistant forms thereof, such as amplification to confirm for presence of MRSA or other target disease

摘要翻译： 用于检测金黄色葡萄球菌（SA）和其临床重要的抗生素抗性形式的方法和组合物，包括耐甲氧西林金黄色葡萄球菌（MRSA），耐万古霉素金黄色葡萄球菌（VRSA），金霉素耐金黄葡萄球菌（Staphylococcus aureus） （mupSA）等来自样本群体中的个体披露还公开了经由固有和快速的SA细胞破碎方法进行细菌抽样和分析的成本有效的方法和试剂盒，其后为直接PCR，绕过需要，费用和污染＆pgr ;与DNA分离方法相关的sks应用这些与SA流行率分析相结合的改进方法，以消除人体中不携带SA（SA阴性）的大约70％的样品，然后进行第二次更昂贵的测试 抗生素抗性形式，例如扩增以证实存在MRSA或其他目标疾病 se