Integrated apparatus for performing nucleic acid extraction and diagnostic testing on multiple biological samples
The technology described herein generally relates to systems for extracting polynucleotides from multiple samples, particularly from biological samples, and additionally to systems that subsequently amplify and detect the extracted polynucleotides. The technology more particularly relates to microfluidic systems that carry out PCR on multiple samples of nucleotides of interest within microfluidic channels, and detect those nucleotides.
1. A preparatory apparatus configured to extract nucleic acid simultaneously from a number of nucleic-acid containing samples comprising:
one or more racks, each rack comprising a plurality of lanes, wherein each lane comprises a first location adjacent to and spatially separate from a second location, the first location comprising a slot shaped and sized to receive a holder, the second location comprising an aperture shaped and sized to receive a sample tube;
a plurality of sample tubes and a corresponding number of holders removably mounted in one of the one or more racks, wherein each holder is removably mounted in the rack adjacent to and spatially separate from its corresponding sample tube, wherein each sample tube is removably mounted in the rack, and wherein each sample tube is configured to accept one of the number of nucleic-acid containing samples, and wherein each holder comprises a process chamber, a waste chamber, one or more pipette tips, and one or more receptacles, wherein the one or more receptacles contain sufficient quantities of one or more reagents for carrying out extraction of nucleic acid from one of the number of nucleic-acid containing sample;
a magnetic separator positioned to apply a magnetic force to the process chamber of each holder when each holder is mounted in the rack;
a heater assembly configured to independently heat each of the process chamber of each holder when each holder is mounted in the rack; and
a liquid dispenser configured to carry out fluid transfer operations between the plurality of sample tubes and the corresponding number of holders.
2. The apparatus of claim 1 , further comprising:
a sample identification verifier configured to check an identity of each of the number of nucleic-acid containing samples.
3. The apparatus of claim 2 , wherein the sample identification verifier is selected from the group consisting of: an optical character reader, a bar code reader, and a radio frequency tag reader.
4. The apparatus of claim 1 , wherein the liquid dispenser has 4 heads, wherein each head is configured to accept a pipette tip from the one or more pipette tips in a holder.
5. The apparatus of claim 4 , wherein no two heads accept pipette tips from the same holder.
6. The apparatus of claim 1 , wherein the liquid dispenser is attached to a gantry having 3 degrees of translational freedom.
7. The apparatus of claim 1 , further comprising:
electronic circuitry configured to control operation of: the liquid dispenser; the heater assembly; and the magnetic separator.
8. The apparatus of claim 7 , wherein the electronic circuitry controls the motion of the liquid dispenser.
9. The apparatus of claim 7 , wherein the electronic circuitry is configured to cause the heater assembly to apply heat independently to the process chambers.
10. The apparatus of claim 7 , wherein the electronic circuitry is configured to cause the magnetic separator to move repetitively relative to the process chamber of each holder.
11. The apparatus of claim 1 , wherein the holders are suitable for a single use and are disposable.
12. The apparatus of claim 1 , wherein the magnetic separator is configured to move repetitively relative to the process chamber of each holder.
13. The apparatus of claim 1 , having two racks.
14. The apparatus of claim 1 , wherein the number of nucleic-acid containing samples is 12.
15. The apparatus of claim 1 , further comprising a second module configured to receive and to store nucleic acid extracted from the number of nucleic-acid containing samples, wherein the second module has separate locations for nucleic acid extracted from each of the nucleic-acid containing samples.
16. The apparatus of claim 1 , further comprising a second module configured to receive and to store nucleic acid extracted from the number of nucleic-acid containing samples, wherein the second module comprises a separate microfluidic PCR cartridge for nucleic acid extracted from each of the number of nucleic-acid containing samples.
17. The apparatus of claim 16 , wherein the microfluidic PCR cartridge contains lyophilized PCR reagents.
18. The apparatus of claim 16 , wherein the microfluidic PCR cartridge contains a first lyophilized PCR reagents suitable for detecting a first analyte, and a second lyophilized PCR reagents suitable for detecting a second analyte.
19. The apparatus of claim 1 , further comprising a second module configured to receive and to store nucleic acid extracted from the number of nucleic-acid containing samples, wherein the second module comprises a Peltier cooler.
20. The apparatus of claim 1 , further comprising a second module configured to receive and to store nucleic acid extracted from the number of nucleic-acid containing samples, wherein the second module has a plurality of areas for receiving nucleic acid extracted from the number of nucleic-acid containing samples.
21. The apparatus of claim 20 , wherein the plurality of areas are configured to be cooled independently of one another.
22. The apparatus of claim 20 , wherein the apparatus comprises more than one rack.
23. The apparatus of claim 1 , wherein each of the holders comprises a pipette tip sheath configured to surround the one or more pipette tips.
24. The apparatus of claim 23 , wherein the pipette sheath is configured to hold 4 pipette tips.
25. The apparatus of claim 1 , wherein each of the holders comprises an identifier.
26. The apparatus of claim 25 , wherein the identifier is a bar-code.
27. The apparatus of claim 1 , wherein each of the holders contains lyophilized sample preparation reagents.
28. The apparatus of claim 1 , wherein one of the one or more reagents comprises: a wash buffer, a release buffer, or a neutralization buffer.
29. The apparatus of claim 1 , wherein the process chamber of each holder has a stellated ridge-pattern on its interior bottom surface.
30. The apparatus of claim 1 , wherein the apparatus is configured to carry out extraction of nucleic acid from the number of nucleic-acid containing samples in less than about 30 minutes.
31. The apparatus of claim 1 , wherein the one or more racks are removable.
32. The apparatus of claim 1 , wherein each rack is configured to accept the number of nucleic-acid containing samples and the corresponding number of holders when disposed outside of the apparatus.
33. The preparatory apparatus of claim 1 , wherein the apparatus is portable or can be situated on a laboratory bench-top.
34. The preparatory apparatus of claim 1 , wherein the number of nucleic-acid containing samples are each independently selected from the group consisting of: saliva, urine, blood, semen, mucus, and spinal fluid.
35. The preparatory apparatus of claim 1 , wherein the liquid dispenser is configured to carry out fluid transfer operation on two or more lanes simultaneously, said fluid transfer operations requiring only two-dimensional movement of the liquid dispenser in each lane.