Automated process for isolating and amplifying a target nucleic acid sequence
원문보기
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
C12P-019/34
C12Q-001/68
C07H-021/02
C07H-021/04
C07H-021/00
출원번호
US-0985064
(2001-11-01)
발명자
/ 주소
Ammann, Kelly G.
Burns, Ralph E.
Hansberry, Ernest V.
Horner, Glenn A.
Jakub, Cheryl A.
Kling, John E.
Nieglos, Donald J.
Schneider, Robert E.
Smith, Robert J.
출원인 / 주소
Gen-Probe Incorporated
인용정보
피인용 횟수 :
106인용 특허 :
206
초록▼
An automated analyzer for performing multiple diagnostic assays simultaneously includes multiple stations, or modules, in which discrete aspects of the assay are performed on fluid samples contained in reaction receptacles. The analyzer includes stations for automatically preparing a specimen sample
An automated analyzer for performing multiple diagnostic assays simultaneously includes multiple stations, or modules, in which discrete aspects of the assay are performed on fluid samples contained in reaction receptacles. The analyzer includes stations for automatically preparing a specimen sample, incubating the sample at prescribed temperatures for prescribed periods, preforming an analyte isolation procedure, and ascertaining the presence of a target analyte. An automated receptacle transporting system moves the reaction receptacles from one station to the next. The analyzer further includes devices for carrying a plurality of specimen tubes and disposable pipette tips in a machine-accessible manner, a device for agitating containers of target capture reagents comprising suspensions of solid support material and for presenting the containers for machine access thereto, and a device for holding containers of reagents in a temperature controlled environment and presenting the containers for machine access thereto. A method for performing an automated diagnostic assay includes an automated process for isolating and amplifying a target analyte. The process is performed by automatically moving each of a plurality of reaction receptacles containing a solid support material and a fluid sample between stations for incubating the contents of the reaction receptacle and for separating the target analyte bound to the solid support from the fluid sample. An amplification reagent is added to the separated analyte after the analyte separation step and before a final incubation step.
대표청구항▼
1. A process for isolating and amplifying a target sequence contained in a target nucleic acid present in a fluid sample, the process comprising the steps of:separating the target nucleic acid from other material present in the fluid sample in a separation station; transporting a reaction receptacle
1. A process for isolating and amplifying a target sequence contained in a target nucleic acid present in a fluid sample, the process comprising the steps of:separating the target nucleic acid from other material present in the fluid sample in a separation station; transporting a reaction receptacle containing the separated target nucleic acid 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 reaction receptacle 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 station.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 hypochiorite, 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 nucleic acid 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 the target nucleic acid is hybridized to a capture probe prior to immobilizing the target nucleic acid 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 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.31. The process of claim 30, wherein said analyzer unit is movable.32. The process of claim 1, wherein the material includes non-target nucleic acid.
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Lair, Gary D.; Nguyen, Thanh N.; Li, Haitao; Li, Florence F.; Knight, Byron J.; Heinz, Robert E.; Macioszek, Jerzy A.; Davis, Christopher B.; Scalese, Robert F., System for performing multi-formatted assays.
Heinz, Robert E.; Newell, Dennis; Opalsky, David; Rhubottom, Jason, Systems and methods for distinguishing optical signals of different modulation frequencies in an optical signal detector.
Heinz, Robert E.; Newell, Dennis; Opalsky, David; Rhubottom, Jason, Systems and methods for distinguishing optical signals of different modulation frequencies in an optical signal detector.
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