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    Method for the detection, identification, enumeration and confirmation of circulating cancer and/or hematologic progenitor cells in whole blood
    1.
    发明授权
    Method for the detection, identification, enumeration and confirmation of circulating cancer and/or hematologic progenitor cells in whole blood 失效
    全血中循环癌症和/或血液祖细胞的检测，鉴定，计数和确认方法

    公开(公告)号：US06197523B1

    公开(公告)日：2001-03-06

    申请号：US08976886

    申请日：1997-11-24

    申请人： David L. Rimm ,  Robert A. Levine ,  Stephen C. Wardlaw ,  Paul Fiedler

    发明人： David L. Rimm ,  Robert A. Levine ,  Stephen C. Wardlaw ,  Paul Fiedler

    IPC分类号： G01N33574

    CPC分类号： G01N15/042 ,  G01N33/56972 ,  G01N33/57484 ,  G01N2015/045 ,  Y10S436/813

    摘要： A method for analyzing blood enables one to isolate, detect, enumerate and confirm under magnification the presence or absence of target cancer cells and/or hematologic progenitor cells which are known to circulate in blood. The analysis is performed in a sample of centrifuged anticoagulated whole blood. The analysis involves both morphometric and epitopic examination of the blood sample while the blood sample is disposed in a centrifuged blood sampling tube. The epitopic analysis of the presence or absence of cancer cells relies on the detection of epitopes which are known to present only on cancer cells; and the epitopic analysis of the presence or absence of hematologic progenitor cells relies on the detection of epitopes which are known to present only on hematologic progenitor cells. The targeted epitopes on the target cell types are epitopes which are also known to be absent on normal circulating blood cells; and the target cancer cell epitopes are epitopes which are known to be absent on target hematologic progenitor cells. Fluorophors with distinct emissions are coupled with antibodies which are directed against the targeted epitopes. The morphometric analysis is performed by staining the cells in the blood sample with an intracellular stain such as acridine orange which highlights the intracellular cell structure. Both the morphometric and epitopic analyses are preferably performed at or near the platelet layer of the expanded buffy coat in the centrifuged blood sample. The morphometric analysis and/or the epitopic analysis may be performed under magnification both visually and/or photometrically.

    摘要翻译： 用于分析血液的方法使得能够在放大下分离，检测，枚举和确认已知在血液中循环的靶癌细胞和/或血液祖细胞的存在或不存在。 在离心的抗凝全血样品中进行分析。 该分析涉及血液样品的形态测定和表征检查，同时将血液样品置于离心取样管中。 癌细胞存在或不存在的代表性分析依赖于已知仅存在癌细胞的表位的检测; 并且血液祖细胞存在或不存在的表征分析依赖于已知仅存在血液祖细胞的表位的检测。 目标细胞类型上的靶向表位是已知在正常循环血细胞上不存在的表位; 并且目标癌细胞表位是已知在靶血液祖细胞上不存在的表位。 具有明显排放的荧光素与针对靶向表位的抗体相结合。 通过用细胞内染色剂如吖啶橙染色血液样品中的细胞进行形态测定分析，突出细胞内细胞结构。 形态测定和表征分析优选在离心血液样品中在扩张的血沉棕黄层的血小板层或其附近进行。 形态测定分析和/或表征分析可以在视觉上和/或光度下放大进行。

    Method for the detection, identification, enumeration and confirmation of virally infected cells and other epitopically defined cells in whole blood
    2.
    发明授权
    Method for the detection, identification, enumeration and confirmation of virally infected cells and other epitopically defined cells in whole blood 失效

    公开(公告)号：US07129056B2

    公开(公告)日：2006-10-31

    申请号：US11155856

    申请日：2005-06-20

    申请人： David L. Rimm ,  Paul Fiedler ,  Robert A. Levine ,  Stephen C. Wardlaw

    发明人： David L. Rimm ,  Paul Fiedler ,  Robert A. Levine ,  Stephen C. Wardlaw

    IPC分类号： G01N33/53

    CPC分类号： G01N33/56972 ,  C07K16/109 ,  G01N15/042 ,  G01N33/57484 ,  G01N33/6878 ,  G01N2015/045 ,  Y10S436/81

    摘要： A method for analyzing blood enables one to isolate, detect, enumerate and confirm under magnification the presence of target cells which have expressed surface epitopes that indicate intracellular infection by various viruses or other infectious agents, and also cells which have expressed surface epitopes that indicate the presence of non-infectious medical conditions. The analysis involves the examination of cells in the blood sample for the presence or absence of particular surface epitopes while the blood sample is disposed in a centrifuged blood sampling container. The epitopic analysis for the presence or absence of infected cells, or cells which indicate the presence of non-infectious medical conditions relies on the detection of known target expressed epitopes. The target epitopes on the target cell types are epitopes which are also known to be absent on normal circulating cells in the blood. Fluorophores or other labels with distinct wavelength emissions are coupled with specific binding agents such as lectins, antibodies, aptamers, or the like, which are directed against the target expressed epitopes. The epitopic analyses may be performed in or near the expanded buffy coat layer in the centrifuged blood sample. The epitopic analysis may be performed under magnification either visually and/or photometrically. The blood sampling container is sized to hold between about 1 and about 20 ml, preferably about 10 ml of blood, and contains an insert that occupies about 90–98% of the volume of the container bore in the area of the container where the target cells will, if present, be detected. The insert forces the target cells in question to reside in an annular space in the container which is adjacent to the circumference of the container bore. The entire analysis can be performed in a relatively short period of time which is typically a matter of minutes to single digit hours.

    Method for the detection, identification, enumeration and confirmation of virally infected cells and other epitopically defined cells in whole blood
    3.
    发明授权
    Method for the detection, identification, enumeration and confirmation of virally infected cells and other epitopically defined cells in whole blood 失效

    公开(公告)号：US06911315B2

    公开(公告)日：2005-06-28

    申请号：US10042016

    申请日：2002-01-10

    申请人： David L. Rimm ,  Paul Fiedler ,  Robert A. Levine ,  Stephen C. Wardlaw

    发明人： David L. Rimm ,  Paul Fiedler ,  Robert A. Levine ,  Stephen C. Wardlaw

    IPC分类号： C07K16/10 ,  C12Q1/70 ,  G01N15/04 ,  G01N33/569 ,  G01N33/574 ,  G01N33/68 ,  G01N33/53

    CPC分类号： G01N33/56972 ,  C07K16/109 ,  G01N15/042 ,  G01N33/57484 ,  G01N33/6878 ,  G01N2015/045 ,  Y10S436/81

    摘要： A method for analyzing blood enables one to isolate, detect, enumerate and confirm under magnification the presence of target cells which have expressed surface epitopes that indicate intracellular infection by various viruses or other infectious agents, and also cells which have expressed surface epitopes that indicate the presence of non-infectious medical conditions. The analysis involves the examination of cells in the blood sample for the presence or absence of particular surface epitopes while the blood sample is disposed in a centrifuged blood sampling container. The epitopic analysis for the presence or absence of infected cells, or cells which indicate the presence of non-infectious medical conditions relies on the detection of known target expressed epitopes. The target epitopes on the target cell types are epitopes which are also known to be absent on normal circulating cells in the blood. Fluorophores or other labels with distinct wavelength emissions are coupled with specific binding agents such as lectins, antibodies, aptamers, or the like, which are directed against the target expressed epitopes. The epitopic analyses may be performed in or near the expanded buffy coat layer in the centrifuged blood sample. The epitopic analysis may be performed under magnification either visually and/or photometrically. The blood sampling container is sized to hold between about 1 and about 20 ml, preferably about 10 ml of blood, and contains an insert that occupies about 90-98% of the volume of the container bore in the area of the container where the target cells will, if present, be detected. The insert forces the target cells in question to reside in an annular space in the container which is adjacent to the circumference of the container bore. The entire analysis can be performed in a relatively short period of time which is typically a matter of minutes to single digit hours.

    Method for assaying whole blood for the presence or absence of circulating cancer or other target cell fragments
    4.
    发明授权
    Method for assaying whole blood for the presence or absence of circulating cancer or other target cell fragments 失效
    用于测定全血存在或不存在循环癌或其它靶细胞碎片的方法

    公开(公告)号：US06670197B2

    公开(公告)日：2003-12-30

    申请号：US09800344

    申请日：2001-03-05

    申请人： David L. Rimm ,  Stephen C. Wardlaw ,  Robert A. Levine ,  Paul Fiedler

    发明人： David L. Rimm ,  Stephen C. Wardlaw ,  Robert A. Levine ,  Paul Fiedler

    IPC分类号： G01N33543

    CPC分类号： G01N15/042 ,  G01N33/56972 ,  G01N33/57484 ,  G01N2015/045

    摘要： This method for analyzing blood enables one to isolate, detect, enumerate and confirm under magnification the presence or absence of fragments of target analyte cancer cells which are circulating in blood. The analysis is performed in a sample of centrifuged anticoagulated whole blood. The analysis of the presence or absence of fragments of cancer cells relies on the detection of external or internal binding sites which are known to be present only in or on tumorous cancer cells. Fluorophors with distinct wavelength emissions are coupled with antibodies, or other binding moieties such as complementary nucleotide sequences, which antibodies are directed against the epithelial cell fragment membrane binding sites, such as internal or external surface epitopes on the cell fragments, or internal binding sites on cell organelles; and which nucleotide sequences are complementary to portions of cell fragment RNA and/or DNA. The labled binding agents are humoric or soluble in the blood sample. The labeled fluorometric binding site-specific materials may be coupled to small plastic beads which have a density or specific gravity that is preferably greater than the specific gravity or density of the red blood cells. The target cell fragments are less dense than the red cells, and typically have the same density or specific gravity as the platelets or white blood cells in the blood sample. Any of the labeled beads which couple with target cell analyte fragments will have a density or specific gravity that is less than the red cells in the blood sample. Thus cell fragment/labeled bead couples will gravitate into an area in the centrifuged blood sample which area is somewhere above the centrifuged red cell layer. The detection of the labeled target analyte/particle couples can be performed in situ in the centrifuged blood sample either visually or photometrically.

    摘要翻译： 用于分析血液的方法使得能够在放大下分离，检测，枚举和确认在血液中循环的靶分析物癌细胞的片段的存在或不存在。 在离心的抗凝全血样品中进行分析。 对癌细胞片段的存在或不存在的分析依赖于已知仅存在于肿瘤细胞中或肿瘤细胞中的外部或内部结合位点的检测。 具有不同波长发射的荧光体与抗体或其它结合部分例如互补核苷酸序列偶联，所述抗体针对上皮细胞片段膜结合位点，例如细胞片段上的内部或外部表面表位或内部结合位点 细胞器 哪些核苷酸序列与细胞片段RNA和/或DNA的部分互补。 标记的粘合剂是幽默的或可溶于血液样品。 标记的荧光结合位点特异性材料可以耦合到具有优选大于红细胞的比重或密度的密度或比重的小塑料珠粒。 靶细胞碎片比红细胞密度低，通常具有与血液样品中的血小板或白细胞相同的密度或比重。 与靶细胞分析物片段偶联的任何标记珠粒将具有小于血液样品中红细胞的密度或比重。 因此，细胞碎片/标记的珠子对将被引入离心的血液样品中的该区域位于离心红细胞层上方的区域中。 标记的目标分析物/颗粒对的检测可以在离心的血液样品中在视觉上或光度下原位进行。

    Evacuated container assembly for analysis of a blood sample for the presence or absence of rare events
    5.
    发明授权
    Evacuated container assembly for analysis of a blood sample for the presence or absence of rare events 失效
    抽真空的容器组件，用于分析血液样本是否存在罕见事件

    公开(公告)号：US06444436B1

    公开(公告)日：2002-09-03

    申请号：US09507635

    申请日：2000-02-22

    申请人： David L. Rimm ,  Paul Fiedler ,  Robert A. Levine ,  Stephen C. Wardlaw

    发明人： David L. Rimm ,  Paul Fiedler ,  Robert A. Levine ,  Stephen C. Wardlaw

    IPC分类号： G01N33574

    CPC分类号： G01N33/57488 ,  Y10S436/81

    摘要： A method for analyzing blood enables one to isolate, detect, enumerate and confirm under magnification the presence or absence of target cancer cells and/or hematologic progenitor cells, or other rare events which are known to circulate in blood. The analysis is performed in a sample of centrifuged anticoagulated whole blood. The analysis involves both morphometric and epitopic examination of the blood sample while the blood sample is disposed in a centrifuged blood sampling container. The epitopic analysis of the presence or absence of cancer cells relies on the detection of epitopes which are known to be present on cancer cells and not on normal circulating blood cells; and the epitopic analysis of the presence or absence of hematologic progenitor cells relies on the detection of epitopes which are known to be present on hematologic progenitor cells and not on normal circulating blood cells. The targeted epitopes on the target cell types are epitopes which are also known to be absent on normal circulating blood cells; and the target cancer cell epitopes are epitopes which are known to be absent on target hematologic progenitor cells. Fluorophors with distinct emissions are coupled with antibodies which are directed against the targeted epitopes. The morphometric analysis is performed by staining the cells in the blood sample with an intracellular stain, such as acridine orange, which highlights the intracellular cell structure. Both the morphometric and epitopic analyses are preferably performed in the expanded buffy coat layer in the centrifuged blood sample. The morphometric analysis and/or the epitopic analysis may be performed under magnification both visually and/or photometrically.

    摘要翻译： 用于分析血液的方法使得能够在放大下分离，检测，枚举和确认靶癌细胞和/或血液祖细胞的存在或不存在，或已知在血液中循环的其它罕见事件。 在离心的抗凝全血样品中进行分析。 该分析涉及血液样品的形态测定和表征检查，同时将血液样品置于离心取样容器中。 癌细胞存在或不存在的表位分析依赖于已知存在于癌细胞而不是正常循环血细胞上的表位的检测; 并且血液祖细胞存在或不存在的表征分析依赖于已知存在于血液祖细胞而不是正常循环血细胞上的表位的检测。 目标细胞类型上的靶向表位是已知在正常循环血细胞上不存在的表位; 并且目标癌细胞表位是已知在靶血液祖细胞上不存在的表位。 具有明显排放的荧光素与针对靶向表位的抗体相结合。 通过用细胞内染色剂如吖啶橙染色血液样品中的细胞进行形态测定分析，其突出显示细胞内细胞结构。 形态测定和表征分析优选在离心血液样品中的膨胀的血沉棕黄层中进行。 形态测定分析和/或表征分析可以在视觉上和/或光度下放大进行。

    Method for in vitro detection of formed elements in biological samples
    6.
    发明授权
    Method for in vitro detection of formed elements in biological samples 失效
    体外检测生物样品中形成元素的方法

    公开(公告)号：US5496704A

    公开(公告)日：1996-03-05

    申请号：US312513

    申请日：1994-09-26

    申请人： Paul Fiedler ,  Robert A. Levine ,  Stephen C. Wardlaw

    发明人： Paul Fiedler ,  Robert A. Levine ,  Stephen C. Wardlaw

    IPC分类号： G01N33/569 ,  G01N33/574

    CPC分类号： G01N33/569 ,  G01N33/56966 ,  G01N33/574 ,  Y10S436/80 ,  Y10S436/805 ,  Y10S436/81 ,  Y10S436/813 ,  Y10S436/824

    摘要： A biological sample such as feces, sputum, cervical tissue, pleural fluids, exudates, cytologic specimens, or the like, is tested for the presence or absence of: ova; parasites; microorganisms; inflammatory, neoplastic tissue cells; or other target materials which are indicative of infestation, disease or infection. The sample is mixed with a buffer fluid and placed in a transparent tube which contains a volume-constricting cylindrical insert for gravimetric separation of components of the sample. The mixture is centrifuged, and the annular space between the insert and tube bore is examined under magnification for the presence of the target materials.

    摘要翻译： 对生物样品如粪便，痰液，宫颈组织，胸膜液，渗出物，细胞学标本等进行测试，以证明是否存在：ova; 寄生虫 微生物 炎性，肿瘤组织细胞; 或指示感染，疾病或感染的其他目标材料。 将样品与缓冲液混合并置于含有体积收缩的圆柱形插入物的透明管中，用于重量分离样品组分。 将混合物离心，并且在放大下检查插入件和管孔之间的环形空间以存在目标材料。