Method for applying thermal energy to a receptacle and detecting an emission signal from the receptacle

Patent metadata

Field	Value
Publication number	US-11840727-B2
Application number	US-202017028367-A
Country	US
Kind code	B2
Filing date	Sep 22, 2020
Priority date	Jul 31, 2012
Publication date	Dec 12, 2023
Grant date	Dec 12, 2023

Abstract

Multiple receptacles containing a reaction mixture are transported to corresponding receptacle wells of a thermally-conductive receptacle holder in thermal communication with a support that is in thermal communication with a heat sink. The heat sink is pre-heated to a temperature above ambient temperature, and the temperature of the receptacle holder is cycled. Optical communication is established between each of the receptacle wells and an excitation signal source and an emission signal detector during cycling, and whether an emission signal is emitted from any of the receptacles is determined as an indication of the presence of a target nucleic acid.

First claim

What is claimed is:

1. A method for determining the presence of a target nucleic acid in a sample, the method comprising the automated steps of:
(a) transporting multiple receptacles to each of a plurality of corresponding receptacle wells of a thermally-conductive receptacle holder, each of the receptacles containing a reaction mixture, wherein the receptacle holder is in thermal communication with a support, and wherein the support is in thermal communication with a heat sink;
(b) pre-heating the heat sink to a temperature above ambient temperature with a thermal device disposed within the heat sink;
(c) after step (b), cycling the temperature of the receptacle holder with a thermal element that is in thermal communication with the support and is separate from the thermal device that pre-heats the heat sink;
(d) establishing optical communication between each of the receptacle wells and an excitation signal source and an emission signal detector during step (c); and
(e) determining whether an emission signal is emitted from any of the receptacles during step (d) as an indication of the presence of a target nucleic acid.

2. The method of claim 1 , wherein the support is disposed on the heat sink.

3. The method of claim 2 , wherein a base of the support is disposed on the heat sink.

4. The method of claim 3 , wherein the receptacle holder is affixed to the support.

5. The method of claim 4 , wherein the receptacle holder is affixed to an upright portion of the support, and wherein the thermal element is situated between the receptacle holder and the upright portion of the support.

6. The method of claim 5 , wherein the thermal element is a thermoelectric device.

7. The method of claim 1 , further comprising the step of closing each of the receptacles with a cap prior to step (a).

8. The method of claim 7 , wherein the cap of each associated receptacle is engaged in a frictional fit with a transport mechanism during step (a).

9. The method of claim 8 , wherein the transport mechanism is a pipettor.

10. The method of claim 9 , further comprising stripping each of the receptacles from the transport mechanism after step (a) and prior to step (b).

11. The method of claim 1 , further comprising, after step (a), exerting a force onto each of the receptacles, thereby seating or securing the receptacles in the corresponding receptacle wells.

12. The method of claim 11 , wherein the force is exerted by a cover movable between an open position and a closed position relative to the receptacle holder, wherein the cover does not obstruct access to the receptacle wells when the cover is in the open position, wherein the cover blocks access to the receptacle wells when the cover is in the closed position, and wherein the cover is configured to exert the force onto each of the receptacles when the cover is in the closed position.

13. The method of claim 1 , wherein the heat sink is pre-heated to a temperature that is at or below a nucleic acid annealing temperature in step (b).

14. The method of claim 13 , wherein the heat sink is pre-heated to a temperature that is above about 20° C. and at or below about 64° C. in step (b).

15. The method of claim 1 , wherein the heat sink is pre-heated to a temperature that is between an annealing temperature and an elongation/extension temperature in step (b).

16. The method of claim 15 , wherein the annealing temperature is in a range of about 50° C. to 64° C. and the elongation/extension temperature is in the range of about 72° C. to 80° C.

17. The method of claim 1 , wherein the heat sink is pre-heated to a temperature that is between an annealing temperature and a melting/denaturation temperature in step (b).

18. The method of claim 17 , wherein the annealing temperature is in a range of about 50° C. to 64° C. and the melting/denaturation temperature is in a range of about 94° C. to 98° C.

19. The method of claim 1 , wherein the heat sink is pre-heated to a temperature that is in a range of about 45° C. to about 50° in step (b).

20. The method of claim 1 , wherein the optical communication is established in step (d) with a plurality of optical fibers, each of the optical fibers extending from a through-hole in a corresponding one of the receptacle wells to the excitation signal source and the emission signal detector.

Assignees

• Gen Probe Inc

Inventors

• Buse David
• Combs David Howard
• Hagen Norbert D
• Opalsky David
• Richardson Bruce
• Prasad Anita
• Moravick Keith
• Moore Tyler
• Knight Byron J

Classifications

• C12Q1/686 Primary

  Polymerase chain reaction [PCR] · CPC title

• B01L7/52 Primary

  with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples · CPC title

• G01N21/6452

  Individual samples arranged in a regular 2D-array, e.g. multiwell plates · CPC title

• B01L9/06

  Test-tube stands; Test-tube holders · CPC title

• B01L2200/141

  Preventing contamination, tampering · CPC title