Kathleen Miller Clark

US Patent:

6660472, Dec 9, 2003

Filed:

Jul 17, 2000

Appl. No.:

09/509006

Inventors:

Lise Santoro - Charbonnieres les Bains, FR
Sophie Lopez - Chazay dAzergues, FR
Bruno Colin - Marcy lEtoile, FR
Corinne Jay - Villeurbanne, FR
Cécile Paris - Marcy lEtoile, FR
Kathleen Ann Clark Dickey - Cardiff CA

Assignee:

Bio Merieux - Marcy lEtoile
Gen-Probe Incorporated - San Diego CA

International Classification:

C12Q 168

US Classification:

435 6, 435 911, 435 912, 436 94, 536 231, 536 253, 536 254

Abstract:

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 m; and the apparent volume of the beads (Vb) and the volume of the liquid sample (Ve) are linked by the relationship Ve=. Vb, with ranging between 1. 4 and 10 when the container is tubular in shape, and is not more than 2.

US Patent:

57862087, Jul 28, 1998

Filed:

Oct 18, 1995

Appl. No.:

8/544560

Inventors:

Kathleen A. Clark - Cardiff by the Sea CA
Daniel L. Kacian - San Diego CA

Assignee:

Gen-Probe Incorporated - San Diego CA

International Classification:

C12N 108
C12N 1500
C12Q 168
C07H 2104

US Classification:

430270

Abstract:

Methods for extracting nucleic acids by heating sample cells at about 80-95 degrees C. in a permeabilization reagent containing a non-ionic detergent and a metal chelating agent. The methods of the invention release large fragments of undegraded nucleic acids without physically disrupting the entire cell wall. The nucleic acids are released into solution without bursting the cells and are suitable for research and testing without further purification. The extraction method described herein is rapid, easy to perform, and applicable to a wide variety of cells, including microorganisms. Clinical samples may be screened for the presence or absence of a microorganism by heating the sample at 80-95 degrees Celsius in the presence of a non-ionic detergent and a metal chelating agent, adding to the sample a nucleic acid probe specific to the selected microorganism, incubating the sample under conditions which allow the probe to hybridize to released nucleic acid and detecting whether any hybridized probe is present. A kit for performing the nucleic acid extraction methods disclosed herein is also described.

US Patent:

58374523, Nov 17, 1998

Filed:

Jan 3, 1997

Appl. No.:

8/778582

Inventors:

Kathleen A. Clark - Cardiff by the Sea CA
Daniel L. Kacian - San Diego CA

Assignee:

Gen-Probe Incorporated - San Diego CA

International Classification:

C12Q 168
C12N 100
C12N 106
B01F 1700

US Classification:

435 6

Abstract:

Methods for extracting nucleic acids by heating sample cells at about 80-95 degrees C. in a permeabilization reagent containing a non-ionic detergent and a metal chelating agent. The methods of the invention release large fragments of undegraded nucleic acids without physically disrupting the entire cell wall. The nucleic acids are released into solution without bursting the cells and are suitable for research and testing without further purification. The extraction method described herein is rapid, easy to perform, and applicable to a wide variety of cells, including microorganisms. Clinical samples may be screened for the presence or absence of a microorganism by heating the sample at 80-95 degrees Celsius in the presence of a non-ionic detergent and a metal chelating agent, adding to the sample a nucleic acid probe specific to the selected microorganism, incubating the sample under conditions which allow the probe to hybridize to released nucleic acid and detecting whether any hybridized probe is present. A kit for performing the nucleic acid extraction methods disclosed herein is also described.