Portable preparation, analysis, and detection apparatus for nucleic acid processing

• US 20070141605A1
• Filed: 11/20/2006
• Published: 06/21/2007
• Est. Priority Date: 11/21/2005
• Status: Abandoned Application

• Current Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Original Assignee: Applera Corporation (Thermo Fisher Scientific Incorporated)
• Inventors: Chiang, Yuh-Min, Greenstein, Michael, Vann, Charles
• Application Number:
  US11/602,149
• Publication Number:
  US 20070141605A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: C12M 47/06 Hydrolysis; Cell lysis; Extraction of intracellular or cell wall material lysis of microorganisms C12N1/06; extracting or separating nucleic acids from biological samples C12N15/1003
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/686 Polymerase chain reaction [PCR]
• CPC Class Codes: G01N 27/447 using electrophoresis

Abstract

The present teachings comprise a device and method for lysing and/or purifying biological sample. The device can comprise a cartridge having a chamber containing a biological sample receiving region, a plurality of electrodes, and one or more sieving matrices. The electrodes can be configured to lyse the biological sample through the production of a pulsed electrical field. The electrodes can also be configured to heat lyse the biological sample. The electrodes can also be configured to electrophoretically move the biological sample through one or more sieving matrices. A portion of the sample can be isolated on a membrane. The portion of the sample isolated on the membrane can be amplified and detected. A portion of the sample can be isolated in a collection area present in the cartridge. The portion of the sample isolated in the collection area can be removed from the cartridge.

Claims

1. A cartridge comprising:

a chamber having a first end and a second end;

a first electrode disposed adjacent the first end;

a second electrode disposed adjacent the second end;

a sample receiving area defined in the chamber between the first electrode and the second electrode;

a nucleic acid capture membrane disposed inside the chamber between the first electrode and the second electrode; and

a sieving matrix disposed inside the chamber between the sample receiving area and the nucleic acid capture membrane;

wherein the first electrode and the second electrode are configured to produce an electric field that extends from the first electrode to the second electrode and the second electrode comprises a portion transparent to visible light.

2. The cartridge of claim 1, further comprising a third electrode and a fourth electrode adapted to electrically contact a buffer disposed in the same receiving space.

3. The cartridge of claim 1, wherein the sieving matrix comprises a micro-porous filter, a frit layer, a bead layer, a fiber composite layer, or a laser drilled membrane.

4. The cartridge of claim 1, further comprising a collection device comprising a cap, wherein the chamber further comprises an opening in fluid communication with the sample receiving area, and the cap is adapted to seal the opening.

5. The cartridge of claim 1, further comprising nucleic acid amplification reagents disposed at least in the capture membrane.

6. The cartridge of claim 1, wherein at least one of the first and the second electrodes is configured to function as a resistive heater.

7. The cartridge of claim 6, further comprising a control unit adapted to use the resistive heater for thermal cycling a sample disposed in the sample receiving area.

8. A system comprising:

the cartridge of claim 1;

and

a cartridge processing device, the cartridge processing device comprising;

a housing, a receptacle formed in the housing and adapted to receive the cartridge, an excitation source adapted to illuminate components in the chamber, a detection device adapted to comprise a field of view of at least a portion of the chamber, and a first electrical contact and a second electrical contact adapted to form electrical communications with the first and second electrodes, respectively.

9. The system of claim 8, wherein the detection device comprises a detection window disposed in the housing.

10. The system of claim 8, wherein the detection device comprises one or more optical filters disposed in the housing, wherein the one or more optical filters are adapted to filter light emitted from the cartridge.

11. The system of claim 8, wherein the detection device comprises one or more lenses disposed in the housing, wherein the one or more lenses are adapted to refract light emitted from the cartridge.

12. The system of claim 8, wherein the cartridge processing device further comprises a power source adapted to provide electrical current to the first and second electrical contacts. wherein the electrical current is configured to form a pulse electric field between the first electrode and the second electrode in a desired pattern.

13. The system of claim 12, further comprising a control unit adapted to control the power supply to generate a direct current between the first electrode and the second electrode, when a buffer is disposed in the cartridge.

14. A cartridge comprising:

a chamber having a first end, a second end, a side wall, and an opening defined in the first end;

a cap disposed in the opening, the cap comprising at least one first electrode;

at least one second electrode disposed at the second end of the chamber;

a first sieving matrix disposed in the chamber between the cap and the at least one second electrode; and

a second sieving matrix disposed in the chamber between the first sieving matrix and the at least one second electrode, the first and second sieving matrixes being spaced from one another to provide a collection area disposed in the chamber between the first sieving matrix and the second sieving matrix.

15. The cartridge of claim 14, wherein the first sieving matrix comprises a micro-porous filter, a frit layer, a bead layer, a fiber composite layer, a cellulose acetate membrane, a porous-polymer membrane, or a laser drilled membrane, and the second sieving matrix comprises agarose, polyethylene, or acrylamide.

16. A method of sample preparation, comprising:

loading a biological sample into a cartridge, the cartridge comprising first and second ends, at least one first electrode at the first end, at least one second electrode at the second end, a first sieving matrix disposed between the at least one first electrode and the at least one second electrode, a second sieving matrix disposed between the first sieving matrix and the at least one second electrode and spaced apart from the first sieving matrix;

electroporating the sample by applying a voltage to the at least one first electrode and the at least one second electrode;

moving polar analytes in the sample into the first sieving matrix by an electrically created motive force; and

collecting a portion of the sample resulting from the electrophoretic moving.

17. The method of claim 16, further comprising lysing the sample by chemical lysing, heating, or sonicating the sample.

18. The method of claim 16, wherein the collecting comprises removing a portion of the sample from the cartridge.

19. The method of claim 16, wherein the detecting comprises visually detecting the emission beams.

20. The method of claim 16, further comprising thermal cycling the biological sample.