Patent
Integrated PCR reactor for cell lysis, nucleic acid isolation and purification, and nucleic acid amplication related applications
| العنوان: | Integrated PCR reactor for cell lysis, nucleic acid isolation and purification, and nucleic acid amplication related applications |
|---|---|
| Patent Number: | 9,409,166 |
| تاريخ النشر: | August 09, 2016 |
| Appl. No: | 12/331508 |
| Application Filed: | December 10, 2008 |
| مستخلص: | Disclosed are integrated devices capable of performing a polymerase chain reaction within a single vessel. Also disclosed are related methods of sample analysis. |
| Inventors: | Mauk, Michael G (Greenville, DE, US); Bau, Haim H (Swarthmore, PA, US); Malamud, Daniel (New York, NY, US); Abrams, William (Merion, PA, US) |
| Assignees: | The Trustees of the University of Pennsylvania (Philadelphia, PA, US) |
| Claim: | 1. A microfluidic chip, comprising: a chamber having a volume and a width, the chamber being at least partially defined by a substrate, the chamber comprising an inlet having an effective cross-sectional diameter in the range of from about 1 to about 10,000 microns, and the chamber comprising an outlet, a nucleic acid concentrator disposed within the chamber, the nucleic acid concentrator being porous, the nucleic acid concentrator spans the cross-section of the chamber such that the nucleic acid concentrator is in fluid communication with both the inlet and the outlet, the inlet and the outlet being configured such that fluid exerted into the chamber enters the chamber through the inlet, contacts the nucleic acid concentrator, and exits the chamber through the exit; the nucleic acid concentrator being capable of preferentially and reversibly binding a nucleic acid and the nucleic acid concentrator comprising at least one lysing reagent; and a polymerase chain reaction reagent disposed within a first heat-sensitive encapsulant disposed within the chamber, wherein the microfluidic chip is configured to concentrate, amplify, and detect nucleic acid in the chamber, and wherein the chip is configured to permit contact between a fluid sample introduced to the chamber and the heat-sensitive encapsulant. |
| Claim: | 2. The microfluidic chip of claim 1 , wherein the microfluidic chip is capable of accommodating all steps of a polymerase chain reaction performed on a cellular sample. |
| Claim: | 3. The microfluidic chip of claim 1 , wherein the inlet comprises a cross-sectional dimension in the range of from about 10 to about 1000 microns. |
| Claim: | 4. The microfluidic chip of claim 1 , wherein the inlet comprises a cross-sectional dimension in the range of from about 50 to about 200 microns. |
| Claim: | 5. The microfluidic chip of claim 1 , wherein the outlet comprises a cross-sectional dimension in the range of from about 1 to about 10,000 microns. |
| Claim: | 6. The microfluidic chip of claim 1 , wherein the outlet comprises a cross-sectional dimension in the range of from about 10 to about 1000 microns. |
| Claim: | 7. The microfluidic chip of claim 1 , wherein the outlet comprises a cross-sectional dimension in the range of from about 50 to about 500 microns. |
| Claim: | 8. The microfluidic chip of claim 1 , wherein the chamber comprises an observation port. |
| Claim: | 9. The microfluidic chip of claim 1 , wherein the substrate comprises a polymer, a metal, a metal oxide, a ceramic, a glass, or any combination thereof. |
| Claim: | 10. The microfluidic chip of claim 1 , wherein the chamber comprises one or more valves capable of modulating fluid flow within the chamber. |
| Claim: | 11. The microfluidic chip of claim 1 , wherein fluid flow across the inlet is modulated by a valve. |
| Claim: | 12. The microfluidic chip of claim 1 , wherein fluid flow across the outlet is modulated by a valve. |
| Claim: | 13. The microfluidic chip of claim 1 , wherein the nucleic acid concentrator comprises a membrane. |
| Claim: | 14. The microfluidic chip of claim 1 , wherein the nucleic acid concentrator comprises silica, alumina, nitrocellulose, an oxide-coated polymer, or any combination thereof. |
| Claim: | 15. The microfluidic chip of claim 1 , wherein a lysing reagent comprises a chaotropic agent, urea, guanidine, or any combination thereof. |
| Claim: | 16. The microfluidic chip of claim 1 , wherein the chamber has disposed within it an enzyme, a nucleic acid, a nucleic acid primer, a biological material, a buffer, a salt, a deoxyribonucleotide, a preservative, or any combination thereof. |
| Claim: | 17. The microfluidic chip of claim 1 , further comprising a heater capable of heating material residing within the chamber. |
| Claim: | 18. The microfluidic chip of claim 17 , wherein the heater comprises a thermoelectric heater. |
| Claim: | 19. The microfluidic chip of claim 1 , further comprising a source of microwave radiation capable of irradiating material residing within the chamber. |
| Claim: | 20. The microfluidic chip of claim 1 , further comprising a source of ultraviolet radiation, infrared radiation, or both, capable of illuminating material residing within the chamber. |
| Claim: | 21. The microfluidic chip of claim 1 , wherein the encapsulant comprises a wax. |
| Claim: | 22. The microfluidic chip of claim 1 , wherein the nucleic acid concentrator comprises a disc. |
| Claim: | 23. The microfluidic chip of claim 1 , wherein the nucleic acid concentrator is disposed so as to divide the chamber into first and second regions, and the encapsulant is enclosed within one region of the chamber. |
| Claim: | 24. The microfluidic chip of claim 1 , wherein the nucleic acid concentrator occupies less than the entire width of the chamber. |
| Claim: | 25. The microfluidic chip of claim 1 , further comprising a second heat-sensitive encapsulant disposed within the chamber, the second heat-sensitive encapsulant having a melting temperature that differs from the melting temperature of the first heat-sensitive encapsulant. |
| Claim: | 26. The microfluidic chip of claim 1 , wherein the chamber is defined entirely by parts that are fixed in position relative to one another. |
| Claim: | 27. The microfluidic chip of claim 26 , wherein the inlet and the outlet are in fluid communication with the first heat sensitive encapsulant such that fluid introduced through the inlet contacts the first heat sensitive encapsulant before flowing to the outlet. |
| Claim: | 28. The microfluidic chip of claim 1 , wherein the inlet and the outlet are in fluid communication with the first heat sensitive encapsulant such that fluid introduced through the inlet contacts the first heat sensitive encapsulant before flowing to the outlet. |
| Claim: | 29. The microfluidic chip of claim 1 , wherein the chamber is defined by at least two planar substrates. |
| Claim: | 30. The microfluidic chip of claim 1 , wherein the inlet and the outlet together define a single axis along which both the inlet and the outlet extend. |
| Patent References Cited: | 5552325 September 1996 Nochumson et al. 5955351 September 1999 Gerdes et al. 6783993 August 2004 Malmquist 2002/0119482 August 2002 Nelson et al. 2005/0202504 September 2005 Anderson et al. WO9600301 January 1996 WO 03/106031 December 2003 |
| Primary Examiner: | Forman, Betty |
| Attorney, Agent or Firm: | Baker & Hostetler LLP |
| رقم الانضمام: | edspgr.09409166 |
| قاعدة البيانات: | USPTO Patent Grants |
| الوصف غير متاح. |