Claims
- 1. A process for isolating and amplifying a target nucleic acid sequence that may be present in a fluid sample, the process comprising the steps of:
separating the target sequence from non-target nucleic acid that may be present in the fluid sample in a separation station; transporting the separated target sequence, if present in the fluid sample, in a reaction receptacle from the separation station to an amplification incubation station comprising one or more temperature-controlled incubators; and incubating the contents of the reaction receptacle, to which one or more amplification reagents have been provided, in the amplification incubation station for a period of time and under conditions sufficient to permit the target sequence to be amplified, wherein the separation and amplification incubation stations are contained within a housing, and wherein each of the separating, transporting and incubating steps is automated.
- 2. The process of claim 1 further comprising providing to the reaction receptacle a probe for a period of time and under conditions sufficient to permit the probe to hybridize to the target sequence or an amplicon thereof.
- 3. The process of claim 2, wherein the probe includes a detectable label.
- 4. The process of claim 3, wherein the label is a fluorescent dye or a chemiluminescent compound.
- 5. The process of claim 2 further comprising detecting the presence or absence of the probe hybridized to the target sequence, or an amplicon thereof, as an indication of the presence or absence of members of a target group of organisms or viruses in the fluid sample, wherein the target group consists of at least one organism or virus.
- 6. The process of claim 5, wherein the detecting step comprises determining the amount of light emitted from the reaction receptacle as an indication of the presence or absence of members of the target group of organisms or viruses in the fluid sample.
- 7. The process of claim 5 further comprising determining the amount of members of the target group of organisms or viruses in the fluid sample.
- 8. The process of claim 5 further comprising transporting the reaction receptacle from the amplification incubation station to a hybridization incubation station within the housing prior to providing the probe to the reaction receptacle, wherein the hybridization incubation station comprises one or more temperature-controlled incubators, and wherein the step of transporting the reaction receptacle from the amplification incubation station to the hybridization incubation station is automated.
- 9. The process of claim 8, wherein the probe is provided to the reaction receptacle after transporting the reaction receptacle from the amplification incubation station to the hybridization incubation station.
- 10. The process of claim 8 further comprising transporting the reaction receptacle from the hybridization incubation station to a detection station within the housing prior to the detecting step, wherein the presence or amount of hybridized probe is determined in the detection station, and wherein the step of transporting the reaction receptacle from the hybridization incubation station to the detection station is automated.
- 11. The process of claim 10, wherein the detection station includes a luminometer for determining the amount of light emitted by the contents of the reaction receptacle.
- 12. The process of claim 1 further comprising raising or lowering the temperature of the contents of the reaction receptacle prior to transporting the reaction receptacle to the amplification incubation station.
- 13. The process of claim 8 further comprising raising or lowering the temperature of the contents of the reaction receptacle prior to transporting the reaction receptacle to at least one of the amplification and hybridization incubation stations.
- 14. The process of claim 10 further comprising providing to the reaction receptacle a deactivating reagent for destroying nucleic acids present in the reaction receptacle.
- 15. The process of claim 14, wherein the deactivating reagent comprises a chemical agent selected from the group consisting of solutions of potassium permanganate, formic acid, solutions of sodium hypochlorite, hydrazine, and dimethyl sulfate.
- 16. The process of claim 14 further comprising transporting the reaction receptacle from the detection station to a deactivation station within the housing prior to providing the deactivating reagent to the reaction receptacle, wherein the step of transporting the reaction receptacle from the detection station to the deactivation station is automated.
- 17. The process of claim 1 further comprising providing to the fluid sample a solid support for a period of time and under conditions sufficient to permit the target sequence to be immobilized, directly or indirectly, on the solid support prior to the separating step.
- 18. The process of claim 17, wherein the solid support comprises a magnetically responsive particle having a polynucleotide bound thereto.
- 19. The process of claim 18, wherein a target nucleic acid containing the target sequence is hybridized to a capture probe prior to immobilizing the target sequence on the solid support.
- 20. The process of claim 18, wherein the fluid sample is subjected to a magnetic field during the separating step.
- 21. The process of claim 20 further comprising providing to the reaction receptacle a probe for a period of time and under conditions sufficient to permit the probe to hybridize to the target sequence or an amplicon thereof.
- 22. The process of claim 21 further comprising detecting the presence or absence of the probe hybridized to the target sequence, or an amplicon thereof, as an indication of the presence or absence of members of a target group of organisms or viruses in the fluid sample, wherein the target group consists of at least one organism or virus.
- 23. The process of claim 22 further comprising transporting the reaction receptacle from the amplification incubation station to a hybridization incubation station within the housing prior to providing the probe to the reaction receptacle, wherein the hybridization incubation station comprises one or more temperature-controlled incubators, and wherein the step of transporting the reaction receptacle from the amplification incubation station to the hybridization incubation station is automated.
- 24. The process of claim 23, wherein the probe is provided to the reaction receptacle after transporting the reaction receptacle from the amplification incubation station to the hybridization incubation station.
- 25. The process of claim 23 further comprising transporting the reaction receptacle from the hybridization incubation station to a detection station within the housing prior to the detecting step, wherein the presence or amount of hybridized probe is determined in the detection station, and wherein the step of transporting the reaction receptacle from the hybridization incubation station to the detection station is automated.
- 26. The process of claim 17 further comprising providing to the reaction receptacle a deactivating reagent for destroying nucleic acids present in the reaction receptacle.
- 27. The process of claim 26 further comprising transporting the reaction receptacle from the detection station to a deactivation station within the housing prior to providing the deactivating reagent to the reaction receptacle, wherein the step of transporting the reaction receptacle from the detection station to the deactivation station is automated.
- 28. The process of claim 1, wherein the transporting step is performed with a receptacle transport mechanism.
- 29. The process of claim 1, wherein the reaction receptacle comprises a plurality of receptacle vessels formed as an integral array, each receptacle vessel containing the same or a different fluid sample for simultaneously separating and amplifying the same or different target sequences which may be present in the fluid samples of the receptacle vessels.
- 30. The process of claim 1, wherein the housing defines a self-contained, stand alone analyzer unit.
Parent Case Info
[0001] This application is a continuation of Ser. No. 09/303,030 filed Apr. 30, 1999, which claims the benefit of International Application No. PCT/US99/09343 filed Apr. 30, 1999, which in turn claims the benefit of U.S. Provisional Application No. 60/083,927 filed May 1, 1998.
Provisional Applications (1)
|
Number |
Date |
Country |
|
60083927 |
May 1998 |
US |
Continuations (1)
|
Number |
Date |
Country |
Parent |
09303030 |
Apr 1999 |
US |
Child |
09985064 |
Nov 2001 |
US |