公开(公告)号：US06660472B1

公开(公告)日：2003-12-09

申请号：US09509006

申请日：2000-07-17

申请人： Lise Santoro ,  Sophie Lopez ,  Bruno Colin ,  Corinne Jay ,  Cécile Paris ,  Kathleen Ann Clark Dickey

发明人： Lise Santoro ,  Sophie Lopez ,  Bruno Colin ,  Corinne Jay ,  Cécile Paris ,  Kathleen Ann Clark Dickey

IPC分类号： C12Q168

CPC分类号： C12N1/066 ,  Y10T436/143333

摘要： The invention concerns a lysis method for a biological sample comprising at least one micro-organism of the bacteria type, for releasing a nucleic material of interest belonging to said micro-organism, which consists in: providing in a container a biological sample in liquid medium; providing in said container at least one particulate material, relatively hard, and substantially inert with respect to the nucleic material; submitting the biological sample and particulate material mixture to a movement. The invention is characterised in that in combination, the selected movement is of the vortex type, and satisfies the following conditions: the particulate material consists of beads with diameter between 90 and 150 &mgr;m; and the apparent volume of the beads (Vb) and the volume of the liquid sample (Ve) are linked by the relationship Ve=&agr;. Vb, with &agr; ranging between 1.4 and 10 when the container is tubular in shape, and &agr; is not more than 2.1 when the container is in the shape of a disk; under such conditions, without any addition of reagent and/or additional process, the method consists in releasing directly into the liquid medium the nucleic material in native state and accessible to any reagent in a subsequent process.

摘要翻译： 本发明涉及包含至少一种细菌类微生物的生物样品的裂解方法，用于释放属于所述微生物的感兴趣的核酸，其包括：在容器中提供液体培养基中的生物样品 ; 在所述容器中提供相对于所述核材料相对较硬且基本上惰性的至少一种颗粒材料; 将生物样品和颗粒物质混合物提交到运动中。 本发明的特征在于，所选择的运动是涡流型的，并且满足以下条件：颗粒材料由直径在90至150μm之间的珠粒组成; 并且珠子（Vb）的表观体积和液体样品（Ve）的体积通过关系Ve =α连接。 Vb，当容器为管状形状时，α范围在1.4和10之间，当容器为盘形时，α不大于2.1; 在这种条件下，没有任何添加试剂和/或另外的方法，该方法包括将天然状态的核酸材料直接释放到液体介质中，并在随后的方法中可用于任何试剂。

公开(公告)号：US06737235B1

公开(公告)日：2004-05-18

申请号：US08945731

申请日：1997-11-10

申请人： Philippe Cros ,  Abdelhamid Elaissari ,  Claude Mabilat ,  Christian Pichot ,  Marc Rodrigue ,  Lise Santoro

发明人： Philippe Cros ,  Abdelhamid Elaissari ,  Claude Mabilat ,  Christian Pichot ,  Marc Rodrigue ,  Lise Santoro

IPC分类号： C07H2104

CPC分类号： C12N15/101 ,  C12N15/1006 ,  C12Q1/6806

摘要： A method for aqueous phase nucleic acid isolation from a sample, comprising a step of nucleic acid adsorption on a particulate substrate, is disclosed. The method comprises an adsorption reagent preparation step (a) that includes a sol consisting of a aqueous continuous phase and a dispersed particulate substrate phase including a functionalized particulate polymer prepared by polymerizing (1) a first water-soluble acrylamide or acrylamide derivative monomer, (2) at least one cross-linking agent and (3) at least one second water-soluble, cationic and functional monomer, said polymer having a predetermined lower critical solubility temperature (LCST) of 25-45° C.; a contact step (b) wherein the adsorption reagent is contacted with the sample containing the nucleic acid; an adsorption step (c) wherein, to carry out the contact step (b), at least one parameter is selected for the reaction medium, said parameters being a pH no higher than 7, an ionic strength no higher than 10−2 M, and a temperature lower than the polymer LCST; a separation step (d) wherein the dispersed phase is separated from the continuous phase, optionally after it has been observed that adsorption has occurred; and a desorption step (e) wherein the nucleic acid is desorbed from the particulate substrate by increasing the ionic strength until an ionic strength higher than 10−2 M is achieved.

摘要翻译： 公开了一种从样品中分离水相核酸的方法，包括在颗粒基质上的核酸吸附步骤。 该方法包括一种吸附剂制备步骤（a），其包括由水性连续相组成的溶胶和包含通过聚合（1）第一水溶性丙烯酰胺或丙烯酰胺衍生物单体制备的官能化颗粒聚合物的分散颗粒基质相（ 2）至少一种交联剂和（3）至少一种第二水溶性阳离子和功能性单体，所述聚合物具有25-45℃的预定的较低临界溶解度温度（LCST）。 接触步骤（b），其中所述吸附剂与含有所述核酸的样品接触; 吸附步骤（c）其中，为了进行接触步骤（b），为反应介质选择至少一个参数，所述参数为不高于7的pH，不高于10 -2的离子强度 M，温度低于聚合物LCST; 分离步骤（d），其中分散相与连续相分离，任选地在已经观察到发生吸附之后; 和解吸步骤（e），其中通过增加离子强度直到达到高于10μM的离子强度，从核粒子脱离。