DEVICE FOR NUCLEIC ACID PREPARATION

• US 20080121591A1
• Filed: 11/28/2007
• Published: 05/29/2008
• Est. Priority Date: 11/29/2006
• Status: Abandoned Application

• Current Assignee: Canon USA, Inc. (Canon Inc.)
• Original Assignee: Canon USA, Inc. (Canon Inc.)
• Inventors: Knight, Ivor T., Dale, Gregory A., Stone, Michele R.
• Application Number:
  US11/946,503
• Publication Number:
  US 20080121591A1
• Time in Patent Office: Days
• US Class Current: 210/748
• 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: Y10T 436/255 including use of a solid sorbent, semipermeable membrane, or liquid extraction

Abstract

A device for nucleic acid preparation includes an outer housing and an inner housing carried within the outer housing and rotatable with respect to the outer housing. The inner housing defines an inner chamber and includes inlet and outlet ports and a nucleic acid extraction substrate within the inner chamber. The outer housing includes a sample inlet port, wash fluid inlet and outlet ports, and elute fluid inlet and outlet ports, and the ports of the inner housing can be selectively aligned with ports of the outer housing by rotating the inner housing with respect to the outer housing. By aligning one or both ports of the inner housing with the appropriate ports of the outer housing, sample material can be introduced into the chamber and wash and elution fluids can be passed through the chamber. Nucleic acid from the sample introduced into the chamber is captured on the extraction substrate and is released from the extraction substrate by sonication prior to being eluted out of the chamber.

Claims

1. A device for nucleic acid preparation comprising:

a first part defining a chamber therein and having first and second ports communicating with said chamber;

a nucleic acid extraction substrate contained within said chamber; and

a second part having a sample input port, first and second wash ports, and first and second elute ports,

wherein said first and second parts are configured in operative relation with each other and are moveable with respect to each other so that (a) one of said first and second ports of said first part can be selectively aligned with said sample input port to permit a sample material to be introduced to said chamber through said sample input channel, (b) said first and second ports of said first part can be selectively aligned with said first and second wash ports to permit a wash medium to be introduced to said chamber through one of said first and second wash ports and removed from said chamber through the other of said first and second wash ports, or (3) said first and second channels of said first part can be selectively aligned with said first and second elute ports to permit an elution medium to be introduced to said chamber through one of said first and second elute ports and removed from said chamber through the other of said first and second elute ports.

2. The device of claim 1, wherein the first part and second part are molded from at least one material selected from the group of materials consisting of polycarbonate, polystyrene, polypropylene, polyethylene, and acrylic.

3. The device of claim 1, wherein said first part includes a side wall at least partially enclosing the chamber, and wherein at least a portion of said side wall comprises a thin membrane constructed and arranged to allow transmission of energy through the membrane.

4. The device of claim 3, wherein said membrane is constructed and arranged to allow transmission of ultrasonic energy through the membrane.

5. The device of claim 4, wherein said membrane has a thickness of 0.01-0.5 mm.

6. The device of claim 5, wherein said membrane has a thickness of about 0.05 mm.

7. The device of claim 1, wherein said nucleic acid extraction substrate comprises a cellulose-based solid substrate impregnated with lysis material which lyses cells and captures nucleic acid in the cellulose fibers.

8. The device of claim 1, wherein said first part is circular in shape, and wherein said second part includes a circular opening for receiving said first part therein, and wherein said first part is configured to be rotatable within the circular opening of said second part.

9. The device of claim 8, wherein the first and second ports of said first part are disposed on opposite sides of said first part and are spaced from each other by about 180°

.

10. The device of claim 9, wherein the first and second wash ports of said second part are disposed on opposite sides of the circular opening and are spaced from each other by about 180°

, and the first and second elute ports of said second part are disposed on opposite sides of the circular opening and are spaced from each other by about 180°

.

11. The device of claim 10, wherein the first and second wash ports and the first and second elute ports of said second part are aligned along mutually orthogonal directions.

12. A method for extracting nucleic acid from a sample within a cartridge comprising a first part defining a chamber therein and having first and second ports communicating with said chamber and a nucleic acid extraction substrate contained within said chamber and a second part having a sample input port, first and second wash ports, and first and second elute ports, wherein said first and second parts are configured in operative relation with each other and are moveable with respect to each other, said method comprising the steps of:

moving the first and second parts with respect to each other to align the first or second port of the first part with the sample input port of the second part;

introducing sample material into the inner chamber through the sample input port and the aligned first or second inner port under conditions that will allow nucleic acid within the sample material to bind to the nucleic acid extraction substrate;

moving the first and second parts with respect to each other to align the first and second ports of the first part with the first and second wash ports, respectively, of the second part;

introducing wash fluid into the inner chamber through the first wash port and the aligned first port;

removing wash fluid from the inner chamber through the second wash port and the aligned second inner port;

applying energy to release nucleic acid from the substrate;

moving the first and second parts with respect to each other to align the first and second ports of the first part with the first and second elute ports, respectively, of the second part;

introducing an elute medium into the inner chamber through the first elute port and the aligned first port; and

removing elute medium from the inner chamber through the second elute port and the aligned second inner port.

13. The method of claim 12, wherein said step of applying energy is applying ultrasonic energy.

14. The method of claim 13, wherein applying ultrasonic energy comprises:

moving the first and second parts with respect to each other to align the first or second port of the first part with the sample input port of the second part;

introducing a probe sonicator into the inner chamber through sample input port and aligned first or second port so that the probe sonicator is in contact with the contents of the inner chamber; and

applying ultrasonic energy from the probe sonicator to the contents of the inner chamber.

15. The method of claim 13, wherein applying ultrasonic energy comprises:

moving the first and second parts with respect to each other so that neither the first nor second port of the first part is aligned with any port of the second part;

positioning a sonicator exterior to the inner chamber adjacent an outer wall of the inner chamber; and

applying ultrasonic energy to the outer wall of the inner chamber in such a manner that sufficient ultrasonic energy is transmitted through the outer wall to release nucleic acid from the substrate.

16. The method of claim 12, wherein the sample material comprises at least one of whole blood, saliva, spinal fluid, amniotic fluid, serum, urine, or fluid from a vaginal or buccal swab.

17. The method of claim 12, wherein the wash fluid comprises water.

18. The method of claim 12, wherein the nucleic acid extraction substrate comprises a cellulose-based solid substrate impregnated with lysis material which lyses cells and captures nucleic acid in the cellulose fibers.

19. The method of claim 12, wherein the ports of the first part are aligned with ports of the second part by rotating the first part with respect to the second part.

20. A genomic isolation chamber comprising:

a housing;

a chamber within the housing;

a first set of channels, wherein the first set of channels is configured to deliver a first medium to the chamber and extract a second medium from the chamber;

a second set of channels, wherein the second set of channels is configured to deliver a third medium to the chamber and extract a fourth medium from the chamber;

an input channel, wherein the input channel is configured to deliver a sample to the chamber; and

an adsorption substrate, wherein the adsorption substrate is configured to adsorb at least a portion of nucleic acid in the sample, and wherein the genomic isolation chamber is configured so that a combination of flowing the first, second, third, and fourth mediums through or from the chamber isolates the adsorbed nucleic acid in the fourth medium.

21. The genomic isolation chamber according to claim 20, wherein the nucleic acid is released from the adsorption substrate by using either sonication from a sonic emitter inserted through the input channel or by noncontact sonication.