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

Patent metadata

Field	Value
Publication number	US-11230733-B2
Application number	US-202017025735-A
Country	US
Kind code	B2
Filing date	Sep 18, 2020
Priority date	Jul 31, 2012
Publication date	Jan 25, 2022
Grant date	Jan 25, 2022

Abstract

Receptacles containing a reaction mixture are transported to receptacle wells of a thermally conductive receptacle holder, and the temperature of the holder is cycled with a thermoelectric device situated between a support and a first side of the holder while a first force is applied onto a second side of the holder. Optical communication between each well and an excitation signal source and an emission signal detector is established to determine whether an emission signal is emitted from any of the receptacles during temperature cycling as an indication of the presence of a target nucleic acid. The thermoelectric device may be situated between an upright portion of the support and the first side of the holder. A second force may be applied onto a top end of each receptacle in the holder by a cover moveable with respect to the receptacles between an open position and a closed position.

First claim

What is claimed is:

1. A method for performing a nucleic acid amplification reaction, 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 wells are aligned in a single row;
(b) cycling the temperature of the receptacle holder with a thermoelectric device situated between a support and a first side of the receptacle holder;
(c) exerting a first force onto a second side of the receptacle holder during the cycling step without applying a force to any of the receptacles disposed in the receptacle wells, the first and second sides being on opposite sides of the receptacle holder;
(d) establishing optical communication between each of the receptacle wells and an excitation signal source and an emission signal detector during step (b); 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 each of the receptacles is closed with a cap prior to step (a), and wherein each of the closed receptacles is transported to a corresponding one of the receptacle wells with a receptacle transport mechanism.

3. The method of claim 2 further comprising, after step (a), stripping each of the closed receptacles from the receptacle transport mechanism.

4. The method of claim 3 , wherein the receptacle transport mechanism is a pipettor.

5. The method of claim 1 , wherein the reaction mixture comprises a PCR master mix.

6. The method of claim 1 , wherein the thermoelectric device is a Peltier device.

7. The method of claim 1 , wherein the support is in thermal communication with a heat sink.

8. The method of claim 7 further comprising, prior to step (b), heating the heat sink to a temperature above an ambient temperature.

9. The method of claim 8 , wherein the temperature is between about 45 degrees Celsius and about 50 degrees Celsius.

10. The method of claim 1 , wherein the support comprises a base portion and an upright portion, and wherein the thermoelectric device is situated between the upright portion of the support and the first side of the receptacle holder.

11. The method of claim 10 , wherein the base portion of the support is disposed on a heat sink.

12. The method of claim 11 further comprising, prior to step (b), heating the heat sink to a temperature above an ambient temperature.

13. The method of claim 12 , wherein the temperature is between about 45 degrees Celsius and about 50 degrees Celsius.

14. The method of claim 1 further comprising exerting a second force onto a top end of each of the receptacles during step (b), thereby seating or securing each of the receptacles within a corresponding one of the receptacle wells.

15. The method of claim 14 , wherein the second force is exerted by a cover moveable between an open position and a closed position, wherein the cover does not obstruct access to the receptacle wells when the cover is in the open position, and wherein the cover blocks access to the receptacle wells when the cover is in the closed position.

16. The method of claim 15 , wherein the step of exerting the second force comprises rotating the cover between the open position and the closed position.

17. The method of claim 16 , wherein the cover comprises a plurality of flexible extensions, each of the flexible extensions being associated with one of the receptacle wells, and wherein the flexible extensions are attached to and extend laterally from a rigid and rotatable member.

18. The method of claim 15 , wherein the optical communication is established with a plurality of optical fibers, each of the optical fibers extending from an associated one of the receptacle wells to the excitation signal source and the emission signal detector, and wherein a first end of each of the optical fibers moves within a through-hole at the bottom center of a corresponding one of the receptacle wells when the cover moves between the open position and the closed position.

19. The method of claim 1 , wherein the optical communication is established 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.

20. The method of claim 19 , wherein a first end of each optical fiber is disposed outside, within, or extending through the through-hole of an associated one of the receptacle wells.

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

• B01L7/52 Primary

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

• B01L2300/0851

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• G01N2021/6482

  Sample cells, cuvettes · CPC title

• B01L2300/043

  Hinged closures · CPC title

• B01L2200/141

  Preventing contamination, tampering · CPC title