Sample processing device for pretreatment and thermal cycling
View Patent ↗Provided herein is a sample processing apparatus including a sample-containing vessel and a thermal cycling apparatus, wherein the vessel has at least 3 fluidically isolated segments that are each thermally conductive, compressible, and expandable. The thermal cycling unit is capable of amplifying extracted nucleic acid and has at least a first processing station and a second processing station, each processing station defining a temperature zone, wherein thermal cycling is performed by alternatively transferring the contents of the sample vessel between the at least two processing stations.
1. A sample processing apparatus, comprising:
(a) a sample-containing vessel comprising:
an opening to receive at least one sample;
a tubule which comprises opposing walls constructed of a flexible material permitting substantial flattening of a selected section of the sample-containing portion;
at least 3 segments within the tubule, the at least 3 segments comprising: a first segment adjacent to the opening and capable of accepting a sample; a second segment adjacent to the first segment and distal to the opening, and containing a nucleic acid extraction agent; and a third segment adjacent to the second segment and distal to the first segment, and containing a PCR reagent; wherein each segment is fluidically isolated at least in part from an adjacent segment by a pressure gate comprising portions of the opposing walls directly bonded to one another, and wherein the pressure gate is opened by application of fluid pressure on a segment that is fluidically isolated in part by the pressure gate and each segment is so thermally conductive as to permit the transfer of thermal energy to and from the contents of each segment, so compressible as to permit opposing wall portions of each segment to touch one another when each segment is compressed, and so expandable as to receive a volume of fluid expelled from another segment when not so compressed; and
a cover for closing the opening; and
(b) a thermal cycling unit capable of amplifying extracted nucleic acid, the thermal cycling unit having at least a first processing station and a second processing station, each processing station defining a temperature zone, wherein thermal cycling is performed by alternatively transferring the contents of the sample vessel between the at least two processing stations.
2. The sample processing apparatus of claim 1 , wherein the vessel has a filter contained therein structured to separate selected components of a sample material from other components of the sample material.
3. The sample processing apparatus of claim 1 , wherein the nucleic acid extraction reagent is a cell lysis reagent.
4. The sample processing apparatus of claim 1 , wherein the PCR reagent comprises at least one nucleotide, an enzyme, a DNA polymerase, a template DNA, an oligonucleotide, a primer, a dye, a marker, a molecular probe, a buffer, and a detection material.
5. The sample processing apparatus of claim 1 , wherein the vessel contains a micro-array bio-chip for analyzing the sample.
6. The sample processing apparatus of claim 1 , further comprising:
an opening to receive at least one sample containing vessel; and
a detection unit capable of detecting a signal from amplified nucleic acid.
7. The sample processing apparatus of claim 1 , wherein the vessel includes a self-sealing injection channel formed therein, the self-sealing injection channel being capable of fluid communication with the sample material in the sample-containing vessel.
8. The sample processing apparatus of claim 1 , wherein the walls comprise at least two layers.
9. The sample processing apparatus of claim 8 , wherein the walls comprise a first layer having a first permeability; and a second layer exterior to the first layer and having a second permeability lesser than the first permeability.
10. The sample processing apparatus of claim 9 , wherein the first layer is constructed of biocompatible material.
11. The sample processing apparatus of claim 1 , wherein at least a portion of the walls are optically transmissive over a selected wavelength range.
12. The sample processing apparatus of claim 1 , wherein the cover comprises a reservoir.
13. The sample processing apparatus of claim 1 , further comprising a sampling instrument attached to the cover.
14. The sample processing apparatus of claim 13 , wherein the sampling instrument is selected from one of a pipette, a needle, a stick, and a tweezer.
15. The sample processing apparatus of claim 1 , wherein the segments form a substantially linear array.
16. The sample processing apparatus of claim 1 , wherein the segments form a contiguous array.
17. The sample processing apparatus of claim 1 , wherein said sample containing vessel further comprises a fourth segment adjacent to the third segment and distal to the second segment, wherein the fourth segment contains a PCR reagent.
18. The sample processing apparatus of claim 1 , wherein at least one pressure gate extends substantially across the entire width of sample-containing vessel.
19. The sample processing apparatus of claim 1 , wherein said sample containing vessel further comprises a handling portion coupled to the sample containing vessel, the handling portion having a generally rigid construction to facilitate handling of the sample processing apparatus.