Sample processing device for pretreatment and thermal cycling

• US 8,148,116 B2
• Filed: 05/02/2011
• Issued: 04/03/2012
• Est. Priority Date: 12/29/2000
• Status: Expired due to Term

• Current Assignee: Roche Molecular Systems (Roche Holding AG)
• Original Assignee: CHEN Ching-Too
• Inventors: Chen, Shuqi
• Primary Examiner(s): Beisner, William H
• Assistant Examiner(s): HOBBS, MICHAEL L
• Application Number:
  US13/099,317
• Publication Number:
  US 20110207121A1
• Time in Patent Office: 337 Days
• Field of Search: 435/91.2, 435/287.3, 435/303.1, 435/304.2, 435/809
• US Class Current: 435/91.2
• CPC Class Codes: B01L 2200/027 for microfluidic devices
• CPC Class Codes: B01L 2200/10 Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0832 cylindrical, tube shaped
• CPC Class Codes: B01L 2300/0874 Three dimensional network
• CPC Class Codes: B01L 2300/1822 using Peltier elements
• CPC Class Codes: B01L 2300/1861 using radiation
• CPC Class Codes: B01L 2400/0421 electrophoretic flow
• CPC Class Codes: B01L 2400/0481 squeezing of channels or chambers
• CPC Class Codes: B01L 2400/0655 pinch valves
• CPC Class Codes: B01L 3/5027 by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: B01L 3/5082 Test tubes per se
• CPC Class Codes: B01L 7/52 with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples amplification or hybridisation processes per se C12Q1/68; controlling sequential reactions for synthesis B01J19/0046
• CPC Class Codes: G01N 35/1002 Reagent dispensers
• CPC Class Codes: Y10T 436/2575 Volumetric liquid transfer

Abstract

A sample processing device may include an opening, a sample pretreatment unit, a thermal cycling reaction unit, and a detection unit.

Claims

1. A sample processing device, comprising:

an opening to receive at least one sample vessel;

a sample pretreatment unit positioned along the opening and comprising at least one of a compression member, a stationary member, and an energy transfer element, the sample pretreatment unit capable of extracting nucleic acids from the sample and transferring the extracted nucleic acid to the thermal cycling unit without opening the sample vessel;

a thermal cycling reaction unit positioned along the opening and capable of amplifying the extracted nucleic acid, the thermal cycling reaction unit having at least one driver, a first processing station defining a first temperature zone, and a second processing station defining a second temperature zone;

a control system controlling the thermal cycling reaction unit and programmed to actuate the at least one driver to perform thermal cycling by repeatedly transferring contents of the sample vessel back and forth between the first processing station and the second processing station; and

a detection unit capable of detecting a signal from the amplified nucleic acid without opening the sample vessel.

2. The sample processing device of claim 1, wherein:

the first processing station includes a first compression member coupled to the at least one driver and adapted to compress the sample vessel within the opening and a first energy transfer element for transferring energy to or from vessel contents at the first processing station, and

the second processing station includes a second compression member coupled to the at least one driver and adapted to compress the sample vessel within the opening and a second energy transfer element for transferring energy to or from vessel contents at the second processing station,

wherein compression of the sample vessel by of one of the compression members displaces a substance within the sample vessel between the processing stations.

3. The sample processing device of claim 1, wherein the sample pretreatment unit comprises at least a processing station including a compression member adapted to compress the sample vessel within the opening.

4. The sample processing device of claim 3, wherein the processing station of the sample pretreatment unit includes a stationary member opposing the compression member across the opening of the sample vessel, wherein the compression member compresses the sample vessel against the stationary member within the opening.

5. The sample processing device of claim 1, wherein the sample pretreatment unit comprises at least a processing station including an energy transfer element for transferring energy to or from vessel contents at the processing station.

6. The sample processing device of claim 1, wherein the sample pretreatment unit is proximal to the opening and the thermal cycling reaction unit is distal to the sample pretreatment unit.

7. The sample processing device of claim 1, further comprising the sample vessel, wherein the sample vessel comprises an opening to receive at least one sample and a plurality of segments, each segment separated and fluidically isolated at least in part from an adjacent segment by a pressure gate, and at least one segment containing a reagent.

8. The sample processing device of claim 7, wherein the sample vessel comprises at least two segments, the first segment containing a nucleic acid extraction reagent and the second segment containing a PCR reagent.

9. The sample processing device of claim 1, wherein the sample pretreatment unit extracts nucleic acid by using at least one of a cell lysis reagent, boiling the sample, GITC (Guanidinium isothiocyanate), formamide solubilization, or filters to separate nucleic acid from unwanted cellular debris.

10. The sample processing device of claim 3, wherein the sample pretreatment unit transfers the extracted nucleic acid to the thermal cycling unit without opening the sample vessel by compressing the sample vessel using its compression member to displace the extracted nucleic acid within the sample vessel to the thermal cycling unit.

11. A method of processing a sample, comprising:

adding a sample to a sample vessel comprising an opening to receive at least one sample and a plurality of segments, each segment separated and fluidically isolated at least in part from an adjacent segment by a pressure gate, and at least one segment containing a reagent;

introducing the sample vessel into the sample processing device of claim 1;

extracting a nucleic acid from the a sample using the sample pretreatment unit;

transferring the extracted nucleic acid from the sample pretreatment unit to the thermal cycling reaction unit without opening the sample vessel;

amplifying the extracted nucleic acid in the thermal cycling reaction unit by alternately transferring contents of the sample vessel between the at least first processing station and second processing station; and

detecting a signal from the amplified nucleic acid without opening the sample vessel.

12. The method of claim 11, wherein the sample pretreatment unit comprises at least a processing station including a compression member adapted to compress the sample vessel within the opening;

and transferring comprises compressing the sample vessel using the compression member to displace the extracted nucleic acid within the sample vessel to the thermal cycling unit.

13. The method of claim 11, wherein the sample pretreatment unit comprises at least a processing station including an energy transfer element;

and the method further comprises transferring energy to or from vessel contents at the processing station.

14. The method of claim 11, further comprising heating the sample in the first processing station to a first temperature.

15. The method of claim 14, further comprising heating the sample in the second processing station to a second temperature.

16. The method of claim 15, wherein the first temperature is effective to denature nucleic acid in the sample and the second temperature is one at which nucleic acid annealing and nucleic acid synthesis can occur.

17. The method of claim 11, wherein the first processing station includes a first compression member adapted to compress the sample vessel within the opening, and the second processing station includes a second compression member adapted to compress the sample vessel within the opening;

and the method comprises alternately compressing the sample vessel using one of the compression members to displace a substance within the sample vessel between the processing stations.

18. The method of claim 11, wherein the detection unit monitors the signal in real time.

19. The sample processing device of claim 7, wherein the pressure gate is formed by a bonding of opposing wall portions of the sample vessel to one another, and wherein the pressure gate is opened by application of fluid pressure on a segment that is fluidly isolated in part by a pressure gate.

20. The sample processing device of claim 1, wherein the first processing station comprises a first energy transfer element producing thermal energy in the first temperature zone, the second processing station comprises a second energy transfer element producing thermal energy in the second temperature zone, and the device further comprises a thermal controller controlling the first energy transfer element and the second energy transfer element and programmed to cause the first energy transfer element to adopt a first temperature and to cause the second energy transfer element to adopt a second temperature different from the first temperature.

21. The sample processing device of claim 7, wherein the first temperature is effective to denature nucleic acid in the sample and the second temperature is one at which nucleic acid annealing and nucleic acid synthesis can occur.