C - Chemistry – Metallurgy – 12 – M
Patent
C - Chemistry, Metallurgy
12
M
C12M 1/33 (2006.01) C12M 3/08 (2006.01) C12N 1/06 (2006.01)
Patent
CA 2304846
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 µm; and the apparent volume of the beads (Vb) and the volume of the liquid sample (Ve) are linked by the relationship Ve = .alpha..Vb, with .alpha. ranging between 1.4 and 10 when the container is tubular in shape, and .alpha. 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.
Procédé de lyse d'un échantillon biologique comprenant au moins un micro- organisme de type bactérie, pour libérer un matériel nucléique d'intérêt appartenant audit micro-organisme, selon lequel: on dispose dans un contenant un échantillon biologique en milieu liquide, on dispose dans ledit contenant au moins un matériau particulaire, relativement dur, et substantiellement inerte par rapport au matériel nucléique, on soumet le mélange de l'échantillon biologique et du matériau particulaire à un mouvement, caractérisé en ce que, en combinaison: le mouvement choisi est de type vortex, et répond aux conditions suivantes: le matériau particulaire est constitué de billes de diamètre compris entre 90 et 150 µm, et le volume apparent des billes, Vb, et le volume de l'échantillon liquide, Ve, sont liés par la relation Ve = .alpha..Vb, avec .alpha. compris entre 1,4 et 10 lorsque le contenant est de forme tubulaire, et .alpha. est inférieur ou égal à 2,1 lorsque le contenant est de forme disque, moyennant quoi, sans ajout de réactif et/ou étape opératoire supplémentaire, on libère directement dans le milieu liquide le matériel nucléique à l'état natif et accessible à tout réactif de traitement postérieur.
Clark Dickey Kathleen Ann
Colin Bruno
Jay Corinne
Lopez Sophie
Paris Cecile
Bio Merieux
Gen-Probe Incorporated
Norton Rose Or S.e.n.c.r.l. S.r.l./llp
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