ROTARY THERMOCYCLING APPARATUS

• US 20020127709A1
• Filed: 06/06/2001
• Published: 09/12/2002
• Est. Priority Date: 12/07/1998
• Status: Active Grant

• Current Assignee: Her Majesty The Queen In Right Of Canada (Government of Canada)
• Original Assignee: Her Majesty The Queen In Right Of Canada (Government of Canada)
• Inventors: Wheatcroft, Roger George Laurence
• Granted Patent: US 6,448,066 B1
• Time in Patent Office: Days
• US Class Current: 435/303.1
• 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

Abstract

A method for carrying out sequential reactions, e.g., sequential biochemical reactions, and a rotary thermocycling apparatus especially adapted for conducting such reactions, for example, polymerase chain reactions, are disclosed. The apparatus comprises a plurality of stations, e.g., four stations, for receiving biochemical samples in flat-bottomed containers. Heating means are provided to independently control the temperature of each station, transport means are provided to move the containers from one station to another in a predetermined sequence, and at least one station has a spray unit to spray liquid reagents into a container at that station. The method includes sequentially cycling a sample through predetermined temperature changes and spraying the sample with at least one reagent.

Claims

1. A rotary thermocycling apparatus, especially for biochemical reactions, comprising:

(a) a plurality of stations and especially at least four stations for receiving samples in a flat-bottomed container, each station having a flat heated plate on which said container is placed and having means to independently control said heated plate at a pre-determined temperature;

(b) means to move each said flat-bottomed container from one station to another station in a pre-determined sequence;

(c) at least two of said stations having a heating unit adapted to be lowered over a container located on said station; and

(d) at least one station having a spray unit adapted to spray a liquid reagent(s) into a container located at said one station.

2. The apparatus of claim 1 in which said station of (d) is adapted for removal of a cover plate from said container prior to activation of the spray, and for replacement of the cover plate after said spray has terminated.

3. The apparatus of claim 1 or claim 2 in which the container is a flat-bottomed container of a dimension less than that of the station.

4. The apparatus of any one of claims 1-3 in which the heating unit is comprised of a section with a flat lower surface that is adapted to be lowered into said container close to but not in contact with the sample in said container.

5. The apparatus of claim 4 in which said lower surface of the heating unit is lowered into contact with a cover plate therein.

6. The apparatus of any one of claims 1-5 in which the apparatus has a programmer for controlling at least (i) the temperature at each station, (ii) the dwell time in each station, (iii) the duration and timing of the spray(s), and (iv) the number of sequential cycles for the biochemical reaction.

7. The apparatus of any one of claims 1-6 in which the container is adapted to receive a filter having the sample thereon, and to receive a cover plate over said filter.

8. The apparatus of any one of claims 1-7 in which the apparatus is programmable and automated.

9. The apparatus of any one of claims 1-8 in which the apparatus is adapted to process more than one sample at a time, up to one for each station in the apparatus.

10. The apparatus of any one of claims 1-9 in which there is an automatic loader for placing containers into the apparatus and for removing containers therefrom.

11. The apparatus of any one of claims 1-10 in which the heating units are on pistons that are lowered into the containers.

12. The apparatus of any one of claims 1-11 in which the station with the spray unit is followed in sequence by the two stations having heating units.

14. The method of claim 13 in which the biochemical sample is located on a filter, a membrane, a microtitre container or microscope slide in said container.

15. The method of claim 13 or claim 14 in which the method is programmable and automated.

13. A method for a sequential reaction especially a sequential biochemical reaction at different temperatures, comprising:

(a) placing a sample in a flat-bottomed container;

(b) sequentially cycling said sample through predetermined changes in temperature by placing said flat-bottomed container on flat heated plates at each said temperature for a predetermined period of time;

(c) optionally spraying said sample with at least one liquid reagent;

(d) controlling at least (i) the temperature at each station, (ii) the dwell time in each station, (iii) the duration and timing of the spray(s), and (iv) the number of sequential cycles for the biochemical reaction.

16. The method of any one of claims 13-15 in which the sample is subjected to a pre-treatment prior to the method for the sequential reaction.

17. The method of any one of claims 13-16 in which a spacer is placed on the filter and a cover plate is placed on the spacer.

18. The method of any one of claims 13-17 in which the reaction is a polymerase chain reaction.

19. The method of any one of claims 13-17 in which the reaction is for detection of specific DNA sequences.

20. The method of any one of claims 13-17 in which the reaction is for detection of bacteria.

21. The method of any one of claims 13-17 in which the reaction is for enumeration of a distinct type of bacterium or species.

22. The method of any one of claims 13-21 in which the sample is subsequently subjected to a photochemical detection process to detect product of the reaction.

23. The method of claim 22 in which the photochemical detection process comprises fluorescence.

24. The method of claim 22 or claim 23 in which there is electronic recording thereof.

25. The method of claim 24 in which the electronic recording comprises using a video camera.

29. The method of claim 28 in which the sample is on a filter having an underpad of absorbed salts.

26. A rotary thermocycling apparatus especially for biochemical reactions, comprising:

(a) a plurality of stations for heating biochemical samples in a flat-bottomed container at predetermined temperatures;

(b) means to move each said flat-bottomed container from one station to another station in a pre-determined sequence; and

(c) at least one station having a spray unit adapted to spray liquid reagent(s) into a container located at said one station.

27. A method for a sequential biochemical reaction at different temperatures, comprising placing a biochemical sample in a container and sequentially cycling said biochemical sample through predetermined changes in temperature by heating said container on a sequence of flat heated plates for a predetermined period of time.

28. A method for a sequential biochemical reaction at different temperatures, comprising placing a biochemical sample on a filter or membrane and sequentially cycling said biochemical sample through predetermined changes in temperature, at least one step in the sequence involving spraying the sample with at least one liquid reagent.